Bicyclic heterocycle compounds and their uses in therapy

ABSTRACT

The invention relates to new bicyclic heterocycle compounds, to pharmaceutical compositions comprising said compounds and to the use of said compounds in the treatment of diseases, e.g. cancer.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.15/675,052, filed on Aug. 11, 2017 (and published as US 2018-0065959A1), which is a continuation of U.S. patent application Ser. No.15/105,360, filed on Jun. 16, 2016 (and published as US 2017-0029419A1), which is a § 371 National Phase Application based on InternationalApplication No. PCT/GB2014/053778, filed on Dec. 19, 2014 and publishedas WO 2015/092420 on Jun. 25, 2015, which claims priority to GreatBritain Patent Application No. 1406986.8, filed on Apr. 17, 2014 andGreat Britain Patent Application No. 1322755.8, filed on Dec. 20, 2013.The entire contents of US 2018-0065959 A1 are hereby incorporated hereinby reference.

FIELD OF THE INVENTION

The invention relates to new bicyclic heterocycle compounds, topharmaceutical compositions comprising said compounds and to the use ofsaid compounds in the treatment of diseases, e.g. cancer.

BACKGROUND OF THE INVENTION

IAP Family

The family of inhibitor of apoptosis (IAP) proteins comprises 8 members,XIAP, cIAP1, cIAP2, NAIP, ILP2, ML-IAP, survivin and BRUCE (also knownas apollon). Members of the IAP family have been shown to inhibitprogrammed cell death through their ability to directly inhibit membersof the caspase family of apoptotic enzymes, although the precise role ofall 8 members is yet to be fully defined. The common structural featureof all IAP family members is a ˜70 amino acid zinc-binding fold termedthe baculovirus IAP repeat (BIR) domain, which is present in one tothree copies.

Many interactions between IAPs and other proteins are mediated via asurface groove on the BIR domain. BIR domains may be classified by theirpeptide-binding specificity. There are three types of BIR domains; typeIII domains (capable of binding caspase (and caspase-like) peptides witha specificity for proline in the third (P3) position (e.g. XIAP BIR3),type II domains (like type III domains but lacking the prolinerequirement e.g. XIAP BIR2) and type I domains (which do not bindcaspases or similar peptides, e.g. XIAP BIR1) (Eckelman et al. CellDeath and Differentiation 2008; 15: 920-928). BIRs are small (˜70 aminoacids) Zn-coordinated domains and a variety of proteins use theirN-terminal to interact with the BIR domains grooves. BIR antagonistsprevent caspases binding to BIRs and hence result in increased caspaseactivity thereby inducing auto-ubiquitination and proteasomaldegradation of IAPs. IAPs are overexpressed in many cancers includingrenal, melanoma, colon, lung, breast, ovarian and prostate cancers (Tammet al., Clin. Cancer Research 2000; 6(5): 1796-803), and have beenimplicated in tumour growth, pathogenesis and resistance to chemo- andradio-therapy (Tamm 2000).

XIAP

XIAP is a 57 kDa protein with three BIR domains, the second and third ofwhich bind caspases and a RING-type zinc finger (E3 ligase). XIAP bindsseveral proteins in addition to caspases, including ligation substratessuch as TAK1 and cofactor TAB1, MURR1 involved in copper homeostasis(Burstein et al., EMBO 2004; 23: 244-254), endogenous inhibitors such assecond mitochondria-derived activator of caspases (SMAC), and those ofless clear function such as MAGE-D1, NRAGE (Jordan et al., J. Biol.Chem. 2001; 276: 39985-39989).

The BIR3 domain binds and inhibits caspase-9, an apical caspase in themitochondrial pathway of caspase activation. A groove on the surface ofthe BIR3 domain interacts with the N-terminus of the small subunit ofcaspase-9, locking capsase-9 in its inactive monomeric form with anincompetent catalytic site (Shiozaki et al., Mol. Cell 2003; 11:519-527).

In addition to caspase-binding, XIAP also inhibits apoptosis throughother mechanisms. XIAP forms a complex with TAK1 kinase and its cofactorTAB1 that leads to activation of JNK and MAPK signal transductionpathways, in turn leading to activation of NF-κB (Sanna et al., Mol CellBiol 2002; 22: 1754-1766). XIAP also activates NF-κB by promoting NF-κBtranslocation to the nucleus and degradation of IκB (Hofer-Warbinek etal., J. Biol. Chem. 2000; 275: 22064-22068, Levkau et al., Circ. Res.2001; 88: 282-290).

Cells transfected with XIAP are able to block programmed cell death inresponse to a variety of apoptotic stimuli (Duckett et al., EMBO 1996;15: 2685-2694, Duckett et al., MCB 1998; 18: 608-615, Bratton, Lewis,Butterworth, Duckett and Cohen, Cell Death and Differentiation 2002; 9:881-892).

XIAP is ubiquitously expressed in all normal tissues, but it ispathologically elevated in many acute and chronic leukaemias, prostate,lung, renal, and other types of tumours (Byrd et al., 2002; Ferreira etal., 2001; Hofmann et al., 2002; Krajewska et al., 2003; Schimmer etal., 2003; Tamm et al., 2000). In de novo acute myeloid leukaemia (AML),XIAP expression correlates with myelomonocytic French-American-British(FAB) subtypes M4/M5 (P<0.05) and expression of monocytic markers in AMLblasts. In addition, XIAP was found to be overexpressed in normalmonocytes but undetectable in granulocytes. In AML, XIAP expression wassignificantly lower in patients with favourable rather than intermediateor poor cytogenetics (n=74; P<0.05) (Tamm et al., Hematol. J. 2004;5(6): 489-95).

Overexpression renders cells resistant to multi-agent therapy and isassociated with poor clinical outcome in disease including AML, renalcancer, melanoma (Tamm et al., Clin. Cancer Research 2000; 6: 1796-1803)and lung cancer (Hofmann et al., J. Cancer Res. Clin. Oncology 2002;128(10): 554-60).

XIAP is translated by a cap-independent mechanism of translationinitiation that is mediated by a unique internal ribosome entry site(IRES) sequence element located in its 5′ untranslated region. Thisallows XIAP mRNA to be actively translated during conditions of cellularstress when the majority of cellular protein synthesis is inhibited.Translational upregulation of XIAP in response to stress increasesresistance to radiation induced cell death (Holcik et al., Oncogene2000; 19: 4174-4177).

XIAP inhibition has been investigated in vitro via several techniquesincluding RNA silencing, gene knockout, peptidic ligand mimetics andsmall molecule antagonists, and has been shown to promote apoptosis as amonotherapy and to sensitise many tumour types to chemotherapy,including bladder (Kunze et al., 2008; 28(4B): 2259-63). XIAP knockoutmice are born at the expected Mendelian frequency, with no obviousphysical or histological defects, and normal life spans (Harlin et al.,Mol. Cell Biol. 2001; 21(10): 3604-3608). This indicates that lackingXIAP activity is not toxic in normal tissues and suggests a therapeuticwindow over tumour cells. Further studies have shown XIAP is a criticaldiscriminator between apoptosis in type 1 and type 2 cells includinghepatocytes and therefore should be used with caution in patients withunderlying liver conditions (Jost et al., Nature, 2009, 460, 1035-1041).It was noted that the cIAP1 and cIAP2 levels are upregulated in the XIAPknockout mouse and may protect from pathology via a compensatorymechanism, suggesting pan-inhibition may be required for functionalknockout. Similarly, cIAP1 and cIAP2 knockout mice are alsoasympotomatic (Conze et al., Mol. Biol. Cell 2005; 25(8): 3348-56).While lack of any one of the IAPs produced no overt phenotype in mice,deletion of cIAP1 with cIAP2 or XIAP resulted in mid embryonic lethality(Moulin, EMBO J., 2012).

Endogenous IAP antagonists such as SMAC have been used to validatemembers of this family as targets for therapeutic agents. SMAC peptideschemosensitise tumour cells, and in combination with platins and TumourNecrosis Factor α-related apoptosis inducing ligand (TRAIL) inxenografts, results in tumour growth delay (Fulda et al., Nat. Med.2002; 808-815; Yang et al., Cancer Res. 2003; 63: 831-837).

A natural product, embellin, was identified as binding at the surfacegroove of the BIR3 domain of XIAP with similar affinity to the naturalSMAC peptide. Embellin induces apoptosis in cell lines in vitro andresults in tumour growth delay in xenografts (Nikolovska-Coleska et al.,J. Med. Chem. 2004; 47(10): 2430-2440; Chitra et al., Chemotherapy 1994;40: 109-113).

XIAP antisense oligonucleotides have been developed as therapeuticagents for solid tumour and haematological malignancies. In vitro theseantisense oligonucleotides have been shown to knockdown proteinexpression levels by ˜70%, induce apoptosis and sensitise cells tochemotherapy and delay tumour growth in vivo. One of these agents,AEG351156, has been studied in clinical trials (Hu et al., Clin. CancerRes. 2003; 9: 2826-2836; Cummings et al., Br. J. Cancer 2005; 92:532-538).

Small molecule antagonists of XIAP developed include peptidomimetics aswell as synthetic agents. The peptidomimetics target the BIR3 domain,mimicking SMAC disruption of caspase-9 binding to XIAP, have showninduction of apoptosis in a variety of tumour cell lines as a singleagent, as well as chemosensitisers and are being further investigatedclinically (Oost et al., J. Med. Chem. 2004; 47: 4417-4426; Sun et al.,Bioorg. Med. Chem. Lett. 2005; 15: 793-797).

Synthetic small molecule antagonists of BIR3 and BIR2 domains alsodemonstrate anti-tumour activity in several different models, includinginduction of apoptosis by annexin-V staining and IC50s of <10 μM againstover one-third of the NC160 cell line panel. XIAP antagonists alsoinduced dose-dependent cell death of primary-cultured leukaemia cells in5 out of 5 chronic lymphocytic leukaemia cell lines and 4 out of 5 acutemyeloid leukaemia cell lines (Schimmer et al., Cancer Cell 2004; 5:25-35; Berezovskaya et al., Cancer Res. 2005; 65(6): 2378-86).

High levels of XIAP protein in tumour cell lines were inverselycorrelated with sensitivity to some anti-cancer drugs, particularlycytarabine and other nucleosides (Tamm et al., Clin. Cancer Research2000; 6: 1796-1803). XIAP inhibition potentiates TRAIL-induced antitumoractivity in two preclinical models of pancreatic cancer in vivo (Vogler2008). Gene expression and transfection studies suggest that theincreased expression of apoptosis suppressor XIAP plays an importantrole in anoikis resistance and in the survival of circulating humanprostate carcinoma cells, thereby promoting metastasis. Small moleculeantagonists were found to be anti-metastatic in these models(Berezovskaya et al., Cancer Res. 2005; 65(6): 2378-86).

XIAP has also been found to be involved in other pathways associatedwith cancer and other diseases and these may also benefit from XIAPtargeted agents. The E3 ligase activity of the RING finger domain ofXIAP is able to bind both to TAB1 and to an upstream BM P receptor (type1), suggesting that XIAP may signal in a TGF-β-mediated pathway(Yamaguchi et al., EMBO 1999; 179-187). Focal adhesion kinase (FAK)overexpression has been shown to result in upregulated XIAP expression(Sonoda et al., J. Biol. Chem. 2000; 275: 16309-16315). E3 ligases areattractive therapeutic targets and molecules which target this activityin other proteins such as MDM2 are being developed (Vassilev et al.,Science 2004; 303: 844-848). Direct or indirect inhibition of the XIAPligase activity may also be useful in the treatment of cancer and otherdiseases. Dysregulated apoptotic signalling, which would result frominhibition of IAP function in controlling programmed cell death, hasalso been implicated in many diseases, including disorders associatedwith cell accumulation (e.g. cancer, autoimmunity, inflammation andrestenosis) or disorders where excessive apoptosis results in cell loss(e.g. stroke, heart failure, neurodegeneration such as Alzheimer'sdisease, Parkinson's disease, Huntington's disease, amyotrophic lateralsclerosis, AIDS, ischaemia (stroke, myocardial infarction) andosteoporosis).

XIAP is an important apoptotic regulator in experimental autoimmuneencephalomyelitis and a potential pharmacological target for treatingautoimmune diseases such as multiple sclerosis (MS) (Moore et al., 2004;203(1): 79-93). Antisense-mediated knockdown of XIAP reverses paralysisin an animal model of MS suggesting that treatments targeting XIAP, andperhaps other IAPs, may have utility in the treatment of MS (Hebb etal., Curr. Drug Disc. Tech. 2008; 5(1): 75-7).

cIAP1, cIAP-2, XIAP and survivin are overexpressed in malignant pleuralmesothelioma and are responsible for a large degree of the resistance ofcultured mesothelioma cells to cisplatin. Levels of circulating TNF-αare significantly higher in mesothelioma patients prior to surgicaltumor debulking compared with those after surgery. TNF-α increases mRNAand protein levels of IAP-1, IAP-2 and XIAP (Gordon et al., 2007). NF-κBupregulation plays an important survival role in mesotheliomas inresponse to the inflammatory effects of exposure to asbestos fibres(Sartore-Bianchi et al., 2007). IAP antagonists have the potential toreverse the pro-survival effect of TNF-α.

The ability of cell lines to upregulate TNF-alpha expressionsufficiently to act in an autocrine fashion and kill the cells, oncecIAP1 & 2 are depleted, is believed to be important for IAP activity(Nature Reviews Cancer (2010), 10(8), 561-74, Gryd-Hansen, M). In vivo,however, certain tumour types are surrounded by a pro-inflammatorycytokine network and hence the tumour cells which, on depletion ofcIAP1/2 are switched towards cell killing by apoptosis, may be triggeredto apoptose by TNF-alpha (or other Death Receptor cytokine agonists)already being produced by surrounding cells in the tumourmicroenvironment, such as tumour-associated macrophages, or indeed bythe tumour cells themselves. Certain tumour types such as breast,ovarian and melanoma display this “inflammatory phenotype” which couldpotentially be targeted by IAP antagonists.

cIAP1 and cIAP2

Cellular IAP (cIAP) 1 and 2 are closely related members of the IAPfamily with three BIR domains, a RING domain and a caspase-recruitment(CARD) domain. A functional nuclear export signal exists within the CARDdomain of cIAP1 which appears to be important for cell differentiation(Plenchette et al., Blood 2004; 104: 2035-2043). The presence of thisCARD domain is unique to cIAP1 and cIAP2 within the IAP family ofproteins. These two genes reside in tandem on chromosome 11q22 and giventheir high degree of similarity are thought to have arisen via geneduplication.

cIAP1, like XIAP and survivin, is widely expressed in tumour cell lines,and has been found to be expressed at high levels in colorectal cancersin particular, as well as lung, ovarian, renal, CNS and breast cancers(Tamm et al., Clin. Cancer Res. 2000; 6: 1796-1803). cIAP2 expression isgenerally more restricted and is thought to be regulated thoughconstitutive ubiquitination and degradation by cIAP1 (Conze et al., Mol.Biol. Cell 2005; 25(8): 3348-56; Mahoney et al., PNAS 2008; 105:11778-11783). Immunohistochemistry and western blot analysis identifiedcIAP1 and cIAP2 as potential oncogenes as both are overexpressed inmultiple lung cancers with or without higher copy numbers (Dia et al.,Human Mol. Genetics 2003; 12(7): 791-801). cIAP1 expression levelpreferentially seems to play an important role in low-stageadenocarcinoma (Hofmann et al., J. Cancer Res. Clin. Oncology 2002;128(10): 554-60).

Increased levels of cIAP1 and cIAP2 and reduced levels of endogenousinhibitors are associated with chemoresistance as has been seen forXIAP. cIAP overexpression has been found to correlate in vitro toresistance to DNA alkylating agents such as carboplatin, cisplatin andtopoisomerase inhibitor VP-16 (Tamm et al., Clin. Cancer Res. 2000; 6:1796-1803). Levels of cIAP1 and survivin were found to be high inthyroid cancer cells after cisplatin and doxorubicin treatment. Cellsresistant to chemotherapy such as taxol showed reduced expression ofSMAC and released minimal amounts of this protein from the mitochondria.Down-regulation of cIAP1 and survivin has been found to increase thecytotoxicity of cisplatin and doxorubicin, whereas overexpression ofSMAC improved the efficacy of taxol. However, silencing of cIAP1 andsurvivin by RNA interference restored sensitivity to doxorubicin andcisplatin (Tirro et al.; Cancer Res. 2006; 66(8): 4263-72).

SMAC mimetics such as LBW242 were originally thought to primarily targetXIAP. However studies have shown that cIAP1 was targeted for degradationby autoubiquitination in cells (Yang et al., J. Biol. Chem. 2004;279(17): 16963-16970) and may have contributed to the apoptotic effectsthat resulted. SiRNA of cIAP1 and Tumour Necrosis Factor (TNF)-alphainduction (or stimulation) were found to combine synergistically andrender cell lines more sensitive (Gaither et al. Cancer Res. 2007; 67(24): 11493-11498).

cIAP1 and cIAP2 have been demonstrated to be critical regulators of theNF-κB signalling pathway which is involved in a diverse range ofbiological processes, particularly in innate and adaptive immunity aswell as in proliferation and survival. NF-κB pathway deregulation isassociated with inflammation and cancers including hepatitis andulcerative colitis, gastritis, hepatocellular carcinoma colorectalcancer and gastric cancers, as well as angiogenesis and metastasis (Shenet al., Apoptosis 2009; 14: 348-363).

On ligand binding, the TNF Receptor (TNF-R) recruits TNFR-associatedDeath Domain (TRADD) and receptor-interacting protein (RIP) 1. TRAF2 andcIAP1/cIAP2 are then recruited to form a large membrane complex. RIP1 isubiquitinated and these polyubiquitin chains serve as a docking site fordownstream kinases, resulting in NF-κB pathway signalling effects (Ea etal., Mol. Cell 2006; 22: 245-257; Wu et al., Nat. Cell Biol. 2006; 8:398-406). The extended roles are complex and yet to be fully defined butcIAP1 and cIAP2 are identified as key components of TNF-alpha mediatedNF-κB signalling regulation as well as constitutive(ligand-independent/classical) NF-κB signalling (Varfolomeev et al.,Cell 2007; 131(4): 669-81). cIAP1 and cIAP2 have been shown to bindTRAF2, an adapter protein that functions in both the classical andalternative NF-κB pathways as well as MAPK pathway signalling pathway(Rothe et al., Cell 2005; 83: 1243-1252). cIAP1 and cIAP2 directlytarget RIP1 for ubiquitination in vitro (Betrand et al., Mol. Cell 2008;30: 689-700).

TNF-alpha regulates many cellular functions, including apoptosis,inflammation, immune response, and cell growth and differentiation(Trace et al., Annu. Rev. Med. 1994; 45: 491-503) and therapeutic IAPantagonists may be of benefit in conditions where these functions areaffected.

Production of TNF-alpha is seen in many malignant tumours, and is one ofthe key drivers of cancer-related inflammation that drives tumourdevelopment and/or progression. cIAPs protect cancer cells from thelethal effects of TNF-alpha.

NAIP

NAIP was the first IAP to be discovered (Roy et al., Cell 1995; 80:167-178). NAIP is unique among the IAPs in that it possesses anucleotide-binding and oligomerisation domain, as well as leucine richrepeats which are similar to those contained in proteins normallyinvolved in innate immunity. There are indications that NAIP may also beover expressed in some cancers including breast and oesophageal cancer(Nemoto et al., Exp. Mol. Pathol. 2004; 76(3): 253-9) as well as MS(Choi et al., J. Korean Med. 2007; 22 Suppl: S17-23; Hebb et al., Mult.Sclerosis 2008; 14(5): 577-94).

ML-IAP

Melanoma inhibitor of apoptosis protein (ML-IAP) contains a single BIRand RING finger motif. ML-IAP is a powerful inhibitor of apoptosisinduced by death receptors and chemotherapeutic agents, probablyfunctioning as a direct inhibitor of downstream effector caspases (Vucicet al., Curr. Biol. 2000; 10(21): 1359-66). ML-IAP is also known asBaculoviral IAP repeat-containing protein 7 (BIRC7), Kidney inhibitor ofapoptosis protein (KIAP), RING finger protein 50 (RNF50) and Livin. TheBIR domain of ML-IAP possesses an evolutionarily conserved fold that isnecessary for anti-apoptotic activity. It has been found that themajority of melanoma cell lines express high levels of ML-IAP incontrast to primary melanocytes, which expressed undetectable levels.These melanoma cells were significantly more resistant to drug-inducedapoptosis. Elevated expression of ML-IAP renders melanoma cellsresistant to apoptotic stimuli and thereby potentially contributes tothe pathogenesis of this malignancy.

ILP-2

ILP-2, also known as BIRC8, has a single BIR domain and a RING domain.ILP-2 is expressed only in testis in normal cells, and binds to caspase9 (Richter et al, Mol. Cell. Biol. 2001; 21: 4292-301).

Survivin

Survivin, also known as BIRCS, inhibits both caspase 3 and caspase 7,but its primary function is mitotic progression regulation, rather thanthe regulation of apoptosis. Survivin promotes formation of microtubulesin the mitotic spindle, counteracting apoptosis during cell cycle.Apoptosis inhibition by survivin is predictive of poor outcome incolorectal cancer (Kawasaki et al., Cancer Res. 1998; 58(22): 5071-5074)and stage III gastric cancer (Song et al., Japanese J. Clin. Oncol.2009; 39(5): 290-296).

BRUCE

BRUCE (BIR repeat-containing ubiquitin-conjugating enzyme) is aperipheral membrane protein in the trans-Golgi network with a single BIRdomain, most similar to that of survivin. BRUCE is inhibited via threemechanisms: (i) SMAC binding, (ii) HtrA2 protease and (iii)caspase-mediated cleavage. In addition, BRUCE acts as a E2/E3 ubiquitinligase via ubiquitin-conjugating (UBC) domain.

SUMMARY OF THE INVENTION

The present invention provides compounds of formula (I). The presentinvention provides compounds which are useful in therapy, in particularin the treatment of cancer. The compounds of formula (I) may beantagonists of the IAP family of proteins (IAP), and especially XIAP,and/or cIAP (such as cIAP1 and/or cIAP2) and may be useful in thetreatment of IAP-mediated conditions.

According to a first aspect of the invention, there is provided acompound of formula (I):

or a tautomeric or a stereochemically isomeric form, a pharmaceuticallyacceptable salt or a solvate thereof;whereinX is CR⁴, N or NR³;wherein

-   -   when X is CR⁴, then U represents nitrogen and R⁶ represents oxo;        or    -   when X is N, then U represents carbon and R⁶ represents        hydroxymethyl or —CH(OR^(x))CH₂OR^(z); or    -   when X is NR³, then U represents carbon and R⁶ represents oxo;        dashed bond (        ) represents a single or double bond wherein at least two of        said dashed bonds represent a double bond;        R¹ and R² independently represent hydrogen or methyl;        R³ represents hydrogen, methyl or —NH₂;        R⁴ represents hydrogen, methyl, hydroxymethyl, —NH₂ or fluorine;        R⁵ represents unsubstituted n-butyl or benzyl substituted on the        phenyl group by one or two fluorines; and        R^(x) and R^(z) independently represent hydrogen or methyl.

In a further aspect of the invention there is provided a compound offormula (I) for use in the prophylaxis or treatment of a disease orcondition as described herein, pharmaceutical compositions comprising acompound of formula (I) and processes for the synthesis of compound offormula (I).

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1: ¹H NMR of Example 39. Sample acquired in DMSO-D₆ and calibratedto the non-deuterated solvent residual of DMSO at δ=2.50 ppm. Containedan internal reference standard (TCNB) present as a singlet at δ=8.5 ppm.

FIG. 2: XRPD of Example 39.

FIG. 3: DSC of Example 39.

FIG. 4: ¹H NMR of Example 40. Sample acquired in DMSO-D₆ and calibratedto the non-deuterated solvent residual of DMSO at δ=2.50 ppm. Containedan internal reference standard (TCNB) present as a singlet at δ=8.5 ppm.

FIG. 5: XRPD of Example 40.

FIG. 6: DSC of Example 40.

FIG. 7: ¹H NMR of Example 41. Sample acquired in DMSO-D₆ and calibratedto the non-deuterated solvent residual of DMSO at 2.50 ppm.

FIG. 8: XRPD of Example 41.

FIG. 9: DSC of Example 41.

FIG. 10: ¹H NMR of Example 42. Sample acquired in DMSO-D₆ and calibratedto the non-deuterated solvent residual of DMSO at 2.50 ppm.

FIG. 11: XRPD of Example 42.

FIG. 12: DSC of Example 42.

FIG. 13: XRPD L-(+)-lactate Form B (diffractogram labelled 1) product ofExample 40, reaction mixture at t=0 h (diffractogram labelled 2), after4 days (diffractogram labelled 3) compared with L-(+)-lactate Form C(diffractogram labelled 4) Example 43.

FIG. 14: XRPD of Example 43 iso-structural with Form B at t=0 h(diffractogram labelled 1), progress of the reaction mixtures(diffractograms labelled 2-6), inter-conversion completed after heatingfor t=5 days to give Form C (diffractogram labelled 7).

FIG. 15: ¹H NMR of Example 43. Sample acquired in DMSO-D₆ and calibratedto the non-deuterated solvent residual of DMSO at δ=2.50 ppm. Containedan internal reference standard (TCNB) present as a singlet at δ=8.5 ppm.

FIG. 16: XRPD of Example 43, (diffractogram labelled 1) overlaid withanhydrous L-(+)-Lactic acid (diffractogram labelled 2).

FIG. 17: DSC of Example 43, (thermogram labelled 1) overlaid withanhydrous L-(+)-Lactic acid (thermogram labelled 2).

DEFINITIONS

Unless the context indicates otherwise, references to formula (I) in allsections of this document (including the uses, methods and other aspectsof the invention) include references to all other sub-formula,sub-groups, preferences, embodiments and examples as defined herein.

By “IAP” we mean any of the IAP family members XIAP, cIAP (cIAP1 and/orcIAP2), NAIP, ILP2, ML-IAP, survivin and/or BRUCE, in particular XIAP,cIAP1, cIAP2, ML-IAP, more particularly XIAP, cIAP1 and/or cIAP2, mostparticularly XIAP and/or cIAP1. In particular we mean the BIR domains ofIAP, in particular the BIR domains of XIAP, cIAP1, or cIAP2.

By “one or more IAP family members” we mean any of the IAP familymembers in particular XIAP, cIAP1 and/or cIAP2, more particularly XIAPand/or cIAP1.

“Potency” is a measure of drug activity expressed in terms of the amountrequired to produce an effect of given intensity. A highly potent drugevokes a larger response at low concentrations. Potency is proportionalto affinity and efficacy. Affinity is the ability of the drug to bind toa receptor. Efficacy is the relationship between receptor occupancy andthe ability to initiate a response at the molecular, cellular, tissue orsystem level.

The term “antagonist” refers to a type of receptor ligand or drug thatblocks or dampens agonist-mediated biological responses. Antagonistshave affinity but no agonistic efficacy for their cognate receptors, andbinding will disrupt the interaction and inhibit the function of anyligand (e.g. endogenous ligands or substrates, an agonist or inverseagonist) at receptors. The antagonism may arise directly or indirectly,and may be mediated by any mechanism and at any physiological level. Anexample of indirect antagonism, would be the indirect antagonism of cIAPas a consequence of ubiquination of cIAP resulting in its degradation.As a result, antagonism of ligands may under different circumstancesmanifest itself in functionally different ways. Antagonists mediatetheir effects by binding to the active site or to allosteric sites onreceptors, or they may interact at unique binding sites not normallyinvolved in the biological regulation of the receptor's activity.Antagonist activity may be reversible or irreversible depending on thelongevity of the antagonist-receptor complex, which, in turn, depends onthe nature of antagonist receptor binding.

The term “treatment” as used herein in the context of treating acondition i.e. state, disorder or disease, pertains generally totreatment and therapy, whether for a human or an animal (e.g. inveterinary applications), in which some desired therapeutic effect isachieved, for example, the inhibition of the progress of the condition,and includes a reduction in the rate of progress, a halt in the rate ofprogress, amelioration of the condition, diminishment or alleviation ofat least one symptom associated or caused by the condition being treatedand cure of the condition. For example, treatment can be diminishment ofone or several symptoms of a disorder or complete eradication of adisorder.

The term “prophylaxis” (i.e. use of a compound as prophylactic measure)as used herein in the context of treating a condition i.e. state,disorder or disease, pertains generally to the prophylaxis orprevention, whether for a human or an animal (e.g. in veterinaryapplications), in which some desired preventative effect is achieved,for example, in preventing occurance of a disease or guarding from adisease. Prophylaxis includes complete and total blocking of allsymptoms of a disorder for an indefinite period of time, the mereslowing of the onset of one or several symptoms of the disease, ormaking the disease less likely to occur.

References to the prophylaxis or treatment of a disease state orcondition such as cancer include within their scope alleviating orreducing the incidence of cancer.

As used herein, the term “mediated”, as used e.g. in conjunction withIAP as described herein (and applied for example to variousphysiological processes, diseases, states, conditions, therapies,treatments or interventions) is intended to operate limitatively so thatthe various processes, diseases, states, conditions, treatments andinterventions to which the term is applied are those in which theprotein plays a biological role. In cases where the term is applied to adisease, state or condition, the biological role played by the proteinmay be direct or indirect and may be necessary and/or sufficient for themanifestation of the symptoms of the disease, state or condition (or itsaetiology or progression). Thus, the protein function (and in particularaberrant levels of function, e.g. over- or under-expression) need notnecessarily be the proximal cause of the disease, state or condition:rather, it is contemplated that the mediated diseases, states orconditions include those having multifactorial aetiologies and complexprogressions in which the protein in question is only partiallyinvolved. In cases where the term is applied to treatment, prophylaxisor intervention, the role played by the protein may be direct orindirect and may be necessary and/or sufficient for the operation of thetreatment, prophylaxis or outcome of the intervention. Thus, a diseasestate or condition mediated by a protein includes the development ofresistance to any particular cancer drug or treatment.

The combinations of the invention may produce a therapeuticallyefficacious effect relative to the therapeutic effect of the individualcompounds/agents when administered separately.

The term ‘efficacious’ includes advantageous effects such as additivity,synergism, reduced side effects, reduced toxicity, increased time todisease progression, increased time of survival, sensitization orresensitization of one agent to another, or improved response rate.Advantageously, an efficacious effect may allow for lower doses of eachor either component to be administered to a patient, thereby decreasingthe toxicity of chemotherapy, whilst producing and/or maintaining thesame therapeutic effect. A “synergistic” effect in the present contextrefers to a therapeutic effect produced by the combination which islarger than the sum of the therapeutic effects of the agents of thecombination when presented individually. An “additive” effect in thepresent context refers to a therapeutic effect produced by thecombination which is larger than the therapeutic effect of any of theagents of the combination when presented individually. The term“response rate” as used herein refers, in the case of a solid tumour, tothe extent of reduction in the size of the tumour at a given time point,for example 12 weeks. Thus, for example, a 50% response rate means areduction in tumour size of 50%. References herein to a “clinicalresponse” refer to response rates of 50% or greater. A “partialresponse” is defined herein as being a response rate of less than 50%.

As used herein, the term “combination”, as applied to two or morecompounds and/or agents, is intended to define material in which the twoor more agents are associated. The terms “combined” and “combining” inthis context are to be interpreted accordingly.

The association of the two or more compounds/agents in a combination maybe physical or non-physical. Examples of physically associated combinedcompounds/agents include:

-   -   compositions (e.g. unitary formulations) comprising the two or        more compounds/agents in admixture (for example within the same        unit dose);    -   compositions comprising material in which the two or more        compounds/agents are chemically/physicochemically linked (for        example by crosslinking, molecular agglomeration or binding to a        common vehicle moiety);    -   compositions comprising material in which the two or more        compounds/agents are chemically/physicochemically co-packaged        (for example, disposed on or within lipid vesicles, particles        (e.g. micro- or nanoparticles) or emulsion droplets);    -   pharmaceutical kits, pharmaceutical packs or patient packs in        which the two or more compounds/agents are co-packaged or        co-presented (e.g. as part of an array of unit doses).

Examples of non-physically associated combined compounds/agents include:

-   -   material (e.g. a non-unitary formulation) comprising at least        one of the two or more compounds/agents together with        instructions for the extemporaneous association of the at least        one compound to form a physical association of the two or more        compounds/agents;    -   material (e.g. a non-unitary formulation) comprising at least        one of the two or more compounds/agents together with        instructions for combination therapy with the two or more        compounds/agents;    -   material comprising at least one of the two or more        compounds/agents together with instructions for administration        to a patient population in which the other(s) of the two or more        compounds/agents have been (or are being) administered;    -   material comprising at least one of the two or more        compounds/agents in an amount or in a form which is specifically        adapted for use in combination with the other(s) of the two or        more compounds/agents.

As used herein, the term “combination therapy” is intended to definetherapies which comprise the use of a combination of two or morecompounds/agents (as defined above). Thus, references to “combinationtherapy”, “combinations” and the use of compounds/agents “incombination” in this application may refer to compounds/agents that areadministered as part of the same overall treatment regimen. As such, theposology of each of the two or more compounds/agents may differ: eachmay be administered at the same time or at different times. It willtherefore be appreciated that the compounds/agents of the combinationmay be administered sequentially (e.g. before or after) orsimultaneously, either in the same pharmaceutical formulation (i.e.together), or in different pharmaceutical formulations (i.e.separately). Simultaneously in the same formulation is as a unitaryformulation whereas simultaneously in different pharmaceuticalformulations is non-unitary. The posologies of each of the two or morecompounds/agents in a combination therapy may also differ with respectto the route of administration.

As used herein, the term “pharmaceutical kit” defines an array of one ormore unit doses of a pharmaceutical composition together with dosingmeans (e.g. measuring device) and/or delivery means (e.g. inhaler orsyringe), optionally all contained within common outer packaging. Inpharmaceutical kits comprising a combination of two or morecompounds/agents, the individual compounds/agents may be unitary ornon-unitary formulations. The unit dose(s) may be contained within ablister pack. The pharmaceutical kit may optionally further compriseinstructions for use.

As used herein, the term “pharmaceutical pack” defines an array of oneor more unit doses of a pharmaceutical composition, optionally containedwithin common outer packaging. In pharmaceutical packs comprising acombination of two or more compounds/agents, the individualcompounds/agents may be unitary or non-unitary formulations. The unitdose(s) may be contained within a blister pack. The pharmaceutical packmay optionally further comprise instructions for use.

The term ‘n-butyl’ as used herein refers to a linear alkyl groupcontaining 4 carbon atoms.

The term ‘oxo’ as used herein refers to the group ═O.

Dashed bond (

) represents a single or double bond as required to complete thevalencies of the atoms being linked by the bond. It will be understoodthat in some instances the bond has aromatic character. Dashed bond (

) represents a single or double bond such that the ring containing X andU contains at least two double bonds.

DETAILED DESCRIPTION OF THE INVENTION

It will be understood from formula (I) that the compounds of theinvention can be represented as follows:

wherein Q represents any of A, B or C below:

In one embodiment Q represents A. In one embodiment Q represents B. Inone embodiment Q represents C.

In one embodiment, X represents CR⁴ or N. In an alternative embodiment,X represents CR⁴ or NR³. In an alternative embodiment, X represents N orNR³. In a further embodiment, X represents CR⁴. In a further alternativeembodiment, X represents N. In a yet further alternative embodiment, Xrepresents NR³.

In one embodiment, one of R¹ and R² represents hydrogen and the otherrepresents methyl, or R¹ and R² both represent hydrogen. In oneembodiment, one of R¹ and R² represents hydrogen and the otherrepresents methyl. In a further embodiment, R¹ represents methyl and R²represents hydrogen. In an alternative embodiment, R¹ representshydrogen and R² represents methyl. In a further alternative embodiment,R¹ and R² both represent hydrogen.

In one embodiment, R³ represents hydrogen or methyl. In an alternativeembodiment, R³ represents hydrogen or —NH₂. In a further alternativeembodiment, R³ represents methyl or —NH₂. In a further embodiment, R³represents hydrogen. In a further alternative embodiment, R³ representsmethyl. In a yet further alternative embodiment, R³ represents —NH₂.

In one embodiment, R⁴ represents hydrogen or methyl. In a furtherembodiment, R⁴ represents hydrogen. In an alternative embodiment, R⁴represents methyl.

In one embodiment, R⁵ represents unsubstituted n-butyl or benzylsubstituted by one or two fluorines on the 2, 3 and/or 4 positions ofthe phenyl group. In one embodiment, R⁵ represents unsubstitutedn-butyl. In an alternative embodiment, R⁵ represents benzyl substitutedon the phenyl group by one or two fluorines. In a further embodiment, R⁵represents benzyl substituted by one or two fluorines on the 2, 3 and/or4 positions of the phenyl group. In a further embodiment, R⁵ representsbenzyl substituted by one fluorine on the 2, 3 or 4 position of thephenyl group (i.e. R⁵ represents 2-fluorobenzyl, 3-fluorobenzyl or4-fluorobenzyl). In a further embodiment, R⁵ represents benzylsubstituted by one fluorine on the 4 position of the phenyl group (i.e.R⁵ represents 4-fluorobenzyl). In a further embodiment, R⁵ representsbenzyl substituted by two fluorines on the 2,3, 3,4 or 2,4 positions ofthe phenyl group (i.e. R⁵ represents 2,3-difluorobenzyl,3,4-difluorobenzyl or 2,4-difluorobenzyl). In a yet further embodiment,R⁵ represents benzyl substituted by two fluorines on the 2,4 positionsof the phenyl group (i.e. R⁵ represents 2,4-difluorobenzyl).

In a further embodiment, R⁵ represents unsubstituted n-butyl,4-fluorophenyl or 2,4-difluorophenyl. In a yet further embodiment, R⁵represents 4-fluorophenyl.

In one embodiment R⁶ represents hydroxymethyl or —CH(OR^(x))CH₂OR^(z).In one embodiment R⁶ represents hydroxymethyl.

In one embodiment R⁶ represents —CH(OR^(x))CH₂OR^(z). In one embodiment,one of R^(x) and R^(z) represents hydrogen and the other representsmethyl or R^(x) and R^(z) both represent hydrogen. In a furtherembodiment, R^(x) represents methyl and R^(z) represents hydrogen. In analternative embodiment, R^(x) represents hydrogen and R^(z) representsmethyl. In a further alternative embodiment, R^(x) and R^(z) bothrepresent hydrogen. In a further embodiment, R^(x) represents hydrogenor methyl and R^(z) represents hydrogen. In a further alternativeembodiment, R^(x) and R^(z) both represent methyl.

In one embodiment, R⁶ represents hydroxymethyl, —CH(OH)CH₂OH,—CH(OMe)CH₂OH or —CH(OH)CH₂OMe. In a further embodiment, R⁶ representshydroxymethyl, —CH(OH)CH₂OH or —CH(OMe)CH₂OH. In a yet furtherembodiment, R⁶ represents hydroxymethyl.

In one embodiment R⁶ represents oxo (i.e. ═O).

Sub-Formulae

In one embodiment the compound of formula (I) is wherein:

X is CR⁴, N or NR³;wherein

-   -   when X is CR⁴, then U represents nitrogen and R⁶ represents oxo;        or    -   when X is N, then U represents carbon and R⁶ represents        hydroxymethyl or —CH(OR^(x))CH₂OR^(z); or    -   when X is NR³, then U represents carbon and R⁶ represents oxo;        dashed bond (        ) represents a single or double bond wherein at least two of        said dashed bonds represent a double bond;        one of R¹ and R² represents hydrogen and the other represents        methyl or R¹ and R² both represent hydrogen;        R³ represents hydrogen, methyl or —NH₂;        R⁴ represents hydrogen or methyl;        R⁵ represents unsubstituted n-butyl or benzyl substituted by one        or two fluorines on the 2, 3 and/or 4 positions of the phenyl        group; and        one of R^(x) and R^(z) represents hydrogen and the other        represents methyl or R^(x) and R^(z) both represent hydrogen.

In a further embodiment the compound of formula (I) is wherein:

X is CR⁴, N or NR³;wherein

-   -   when X is CR⁴, then U represents nitrogen and R⁶ represents oxo;        or    -   when X is N, then U represents carbon and R⁶ represents        hydroxymethyl or —CH(OR^(x))CH₂OR^(z); or    -   when X is NR³, then U represents carbon and R⁶ represents oxo;        dashed bond (        ) represents a single or double bond wherein at least two of        said dashed bonds represent a double bond;        one of R¹ and R² represents hydrogen and the other represents        methyl or R¹ and R² both represent hydrogen;        R³ represents hydrogen, methyl or —NH₂;        R⁴ represents hydrogen or methyl;        R⁵ represents unsubstituted n-butyl, 4-fluorobenzyl or        2,4-fluorobenzyl;        R^(x) represents hydrogen or methyl; and        R^(z) represents hydrogen.

In one embodiment, the compound of formula (I) is a compound of formula(Ia):

or a tautomeric or a stereochemically isomeric form, a pharmaceuticallyacceptable salt or a solvate thereof; wherein R¹, R², R⁴ and R⁵ are asdefined in any of the embodiments.

In one embodiment of the compound of formula (Ia), one of R¹ and R²represents hydrogen and the other represents methyl or R¹ and R² bothrepresent hydrogen. In a further embodiment of the compound of formula(Ia), R¹ represents hydrogen and R² represents methyl or R¹ and R² bothrepresent hydrogen.

In a further embodiment of the compound of formula (Ia), R¹ representsmethyl and R² represents hydrogen. In an alternative embodiment of thecompound of formula (Ia), R¹ represents hydrogen and R² representsmethyl.

In one embodiment of the compound of formula (Ia), R⁴ representshydrogen or methyl.

In one embodiment of the compound of formula (Ia), R⁵ representsunsubstituted n-butyl or benzyl substituted by one or two fluorines onthe 2, 3 and/or 4 positions of the phenyl group. In one embodiment ofthe compound of formula (Ia), R⁵ represents unsubstituted n-butyl. In analternative embodiment of the compound of formula (Ia), R⁵ representsbenzyl substituted on the phenyl group by one or two fluorines. In afurther embodiment of the compound of formula (Ia), R⁵ represents benzylsubstituted by one or two fluorines on the 2, 3 and/or 4 positions ofthe phenyl group. In a further embodiment of the compound of formula(Ia), R⁵ represents benzyl substituted by one fluorine on the 4 positionof the phenyl group (i.e. R⁵ represents 4-fluorobenzyl). In a furtherembodiment of the compound of formula (Ia), R⁵ represents benzylsubstituted by two fluorines on the 2,4 positions of the phenyl group(i.e. R⁵ represents 2,4-difluorobenzyl).

In one embodiment, the compound of formula (I) is a compound of formula(Ib):

or a tautomeric or a stereochemically isomeric form, a pharmaceuticallyacceptable salt or a solvate thereof; wherein R¹, R², R⁵, R⁶, R^(x) andR^(z) are as defined in any of the embodiments. In one embodiment R⁶represents hydroxymethyl or —CH(OR^(x))CH₂OR^(z).

In one embodiment of the compound of formula (Ib), R¹ represents methyland R² represents hydrogen or R¹ and R² both represent hydrogen.

In a further embodiment of the compound of formula (Ib), R¹ and R² bothrepresent hydrogen.

In one embodiment of the compound of formula (Ib), R⁵ representsunsubstituted n-butyl or benzyl substituted by one or two fluorines onthe 2, 3 and/or 4 positions of the phenyl group. In one embodiment ofthe compound of formula (Ib), R⁵ represents unsubstituted n-butyl. In analternative embodiment of the compound of formula (Ib), R⁵ representsbenzyl substituted on the phenyl group by one or two fluorines. In afurther embodiment of the compound of formula (Ib), R⁵ represents benzylsubstituted by one or two fluorines on the 2, 3 and/or 4 positions ofthe phenyl group. In a further embodiment of the compound of formula(Ib), R⁵ represents benzyl substituted by one fluorine on the 2, 3 or 4position of the phenyl group (i.e. R⁵ represents 2-fluorobenzyl,3-fluorobenzyl or 4-fluorobenzyl). In a further embodiment of thecompound of formula (Ib), R⁵ represents benzyl substituted by twofluorines on the 2,4 positions of the phenyl group (i.e. R⁵ represents2,4-difluorobenzyl). In a yet further embodiment of the compound offormula (Ib), R⁵ represents benzyl substituted by one fluorine on the 4position of the phenyl group (i.e. R⁵ represents 4-fluorobenzyl).

In one embodiment of the compound of formula (Ib), R⁶ representshydroxymethyl, —CH(OH)CH₂OH, —CH(OMe)CH₂OH or —CH(OH)CH₂OMe.

In a further embodiment of the compound of formula (Ib), R⁶ representshydroxymethyl, —CH(OH)CH₂OH or —CH(OMe)CH₂OH.

In a yet further embodiment of the compound of formula (Ib), R⁶represents hydroxymethyl.

In one embodiment, the compound of formula (I) is a compound of formula(Ic):

or a tautomeric or a stereochemically isomeric form, a pharmaceuticallyacceptable salt or a solvate thereof; wherein R¹, R², R³ and R⁵ are asdefined in any of the embodiments.

In one embodiment of the compound of formula (Ic), one of R¹ and R²represents hydrogen and the other represents methyl or R¹ and R² bothrepresent hydrogen. In a further embodiment of the compound of formula(Ic), R¹ represents methyl and R² represents hydrogen or R¹ and R² bothrepresent hydrogen.

In one embodiment of the compound of formula (Ic), R³ representshydrogen or methyl. In an alternative embodiment of the compound offormula (Ic), R³ represents hydrogen or —NH₂. In a further alternativeembodiment of the compound of formula (Ic), R³ represents methyl or—NH₂.

In a further embodiment of the compound of formula (Ic), R³ representshydrogen. In a further alternative embodiment of the compound of formula(Ic), R³ represents methyl. In a yet further alternative embodiment ofthe compound of formula (Ic), R³ represents —NH₂.

In one embodiment of the compound of formula (Ic), R⁵ representsunsubstituted n-butyl or benzyl substituted by one or two fluorines onthe 2, 3 and/or 4 positions of the phenyl group. In one embodiment ofthe compound of formula (Ic), R⁵ represents unsubstituted n-butyl. In analternative embodiment of the compound of formula (Ic), R⁵ representsbenzyl substituted on the phenyl group by one or two fluorines. In afurther embodiment of the compound of formula (Ic), R⁵ represents benzylsubstituted by one or two fluorines on the 2, 3 and/or 4 positions ofthe phenyl group. In a further embodiment of the compound of formula(Ic), R⁵ represents benzyl substituted by one fluorine on the 2 or 4position of the phenyl group (i.e. R⁵ represents 2-fluorobenzyl or4-fluorobenzyl). In a further embodiment of the compound of formula(Ic), R⁵ represents benzyl substituted by one fluorine on the 4 positionof the phenyl group (i.e. R⁵ represents 4-fluorobenzyl). In a furtherembodiment of the compound of formula (Ic), R⁵ represents benzylsubstituted by two fluorines on the 2,4 positions of the phenyl group(i.e. R⁵ represents 2,4-difluorobenzyl).

In one embodiment, the compound of formula (I) is a compound of formula(Id):

or a tautomeric or a stereochemically isomeric form, a pharmaceuticallyacceptable salt or a solvate thereof; wherein R⁵ is as defined in any ofthe embodiments.

In one embodiment of the compound of formula (Id), R⁵ representsunsubstituted n-butyl or benzyl substituted by one or two fluorines onthe 2, 3 and/or 4 positions of the phenyl group. In one embodiment ofthe compound of formula (Id), R⁵ represents unsubstituted n-butyl. In analternative embodiment of the compound of formula (Id), R⁵ representsbenzyl substituted on the phenyl group by one or two fluorines. In afurther embodiment of the compound of formula (Id), R⁵ represents benzylsubstituted by one or two fluorines on the 2, 3 and/or 4 positions ofthe phenyl group. In a further embodiment of the compound of formula(Id), R⁵ represents benzyl substituted by one fluorine on the 2, 3 or 4position of the phenyl group (i.e. R⁵ represents 2-fluorobenzyl,3-fluorobenzyl or 4-fluorobenzyl). In a further embodiment of thecompound of formula (Id), R⁵ represents benzyl substituted by twofluorines on the 2,4 positions of the phenyl group (i.e. R⁵ represents2,4-difluorobenzyl). In a yet further embodiment of the compound offormula (Id), R⁵ represents benzyl substituted by one fluorine on the 4position of the phenyl group (i.e. R⁵ represents 4-fluorobenzyl).

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib) or (Ic) wherein R¹ represents methyl and R² representshydrogen.

In one embodiment, the compound of formula (I) is a compound of formula(I), (I), (Ib) or (Ic) wherein R¹ and R² both represent hydrogen.

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib) or (Ic) wherein R¹ represents hydrogen and R² representsmethyl.

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib), (Ic) or (Id) wherein R⁵ represents unsubstitutedn-butyl.

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib), (Ic) or (Id) wherein R⁵ represents benzyl substitutedby one or two fluorines on the 2, 3 and/or 4 positions of the phenylgroup.

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib), (Ic) or (Id) wherein R⁵ represents benzyl substitutedon the phenyl group by one or two fluorines.

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib), (Ic) or (Id) wherein R⁵ represents benzyl substitutedon the phenyl group by two fluorines, e.g. 2,3 disubstituted, 2,4disubstituted, 2,5 disubstituted, 3,5 disubstituted, 2,6 disubstitutedor 3,4 disubstituted.

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib), (Ic) or (Id) wherein R⁵ represents benzyl substitutedby one or two fluorines on the 2, 3 and/or 4 positions of the phenylgroup.

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib), (Ic) or (Id) wherein R⁵ represents benzyl substitutedby one fluorine on the 2, 3 or 4 position of the phenyl group (i.e. R⁵represents 2-fluorobenzyl, 3-fluorobenzyl or 4-fluorobenzyl).

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib), (Ic) or (Id) wherein R⁵ represents benzyl substitutedby two fluorines on the 2,3, 3,4 or 2,4 positions of the phenyl group(i.e. R⁵ represents 2,3-difluorobenzyl, 3,4-difluorobenzyl or2,4-difluorobenzyl).

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib), (Ic) or (Id) wherein R⁵ represents benzyl substitutedby two fluorines on the 2,4 positions of the phenyl group (i.e. R⁵represents 2,4-difluorobenzyl).

In one embodiment, the compound of formula (I) is a compound of formula(I), (Ia), (Ib), (Ic) or (Id) wherein R⁵ represents 2,4-difluorobenzylor 4-fluorobenzyl.

In one embodiment, the invention provides a compound of formula (I)which comprises the free base of a compound of Examples 1-37 or atautomeric or a stereochemically isomeric form, a pharmaceuticallyacceptable salt or a solvate thereof.

In one embodiment, the invention provides a compound of formula (I)which is the free base of a compound of Examples 1-37 or a tautomeric ora stereochemically isomeric form, a pharmaceutically acceptable salt ora solvate thereof.

In one embodiment, the invention provides a compound of formula (I)which comprises a compound of Examples 1-37 or a tautomeric or astereochemically isomeric form or a solvate thereof.

In a further embodiment, the compound is selected from the free base ofExamples 1 to 34 or a tautomeric or a stereochemically isomeric form, apharmaceutically acceptable salt or a solvate thereof.

In a further embodiment, the invention provides a compound of formula(I) which comprises a compound selected from:

-   1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one;-   6-[(4-Fluorophenyl)methyl]-3,3,4-trimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one;-   6-[(2,4-Difluorophenyl)methyl]-3,3-dimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one;-   6-[(2,4-Difluorophenyl)methyl]-3,3,4-trimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one;-   1-[5-((R or    S)-1,2-Dihydroxyethyl)-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl]-2-[(2R,5R)-2-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]ethan-1-one;-   6-[(2,4-Difluorophenyl)methyl]-1-{2-[(2R,5R)-2-{[(2S,5R)-2,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]acetyl}-3,3-dimethyl-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-5-one;-   4-Amino-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one;-   1-{6-[(4-Fluorophenyl)methyl]-5-((R or    S)-2-hydroxy-1-methoxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one;-   4-Amino-6-butyl-1-{2-[(2R,5R)-2-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]acetyl}-3,3-dimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one;-   6-[(2,4-Difluorophenyl)methyl]-3,3,4-trimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-5-one;-   6-Butyl-1-{2-[(2R,5R)-2-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]acetyl}-3,3-dimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one;    and-   6-Butyl-1-{2-[(2R,5R)-2-{[(2S,5R)-2,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]acetyl}-3,3-dimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one;    or a tautomeric or a stereochemically isomeric form, a    pharmaceutically acceptable salt or a solvate thereof.

In a further embodiment, the invention provides a compound selectedfrom:

-   1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one    dihydrochloride (E2);-   6-[(4-Fluorophenyl)methyl]-3,3,4-trimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one    dihydrochloride (E6);-   6-[(2,4-Difluorophenyl)methyl]-3,3-dimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one    dihydrochloride (E8);-   6-[(2,4-Difluorophenyl)methyl]-3,3,4-trimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one    dihydrochloride (E19);-   1-[5-((R or    S)-1,2-Dihydroxyethyl)-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl]-2-[(2R,5R)-2-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]ethan-1-one    dihydrochloride (E21);-   6-[(2,4-Difluorophenyl)methyl]-1-{2-[(2R,5R)-2-{[(2S,5R)-2,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]acetyl}-3,3-dimethyl-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-5-one    dihydrochloride (E22);-   4-Amino-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one    dihydrochloride (E24);-   1-{6-[(4-Fluorophenyl)methyl]-5-((R or    S)-2-hydroxy-1-methoxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one    trihydrochloride (E27);-   4-Amino-6-butyl-1-{2-[(2R,5R)-2-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]acetyl}-3,3-dimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one    dihydrochloride (E30);-   6-[(2,4-Difluorophenyl)methyl]-3,3,4-trimethyl-1-{2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]acetyl}-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-5-one    dihydrochloride (E31);-   6-Butyl-1-{2-[(2R,5R)-2-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]acetyl}-3,3-dimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one    dihydrochloride (E32) and-   6-Butyl-1-{2-[(2R,5R)-2-{[(2S,5R)-2,5-dimethylmorpholin-4-yl]methyl}-5-methylpiperazin-1-yl]acetyl}-3,3-dimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one    dihydrochloride (E37)    or a tautomeric or a stereochemically isomeric form, or a solvate    thereof.

In a further embodiment, the compound is selected from the free base ofExamples 2, 6, 19, 21, 22, 24, 27, 30, 31 and 32, or a tautomeric or astereochemically isomeric form, a pharmaceutically acceptable salt or asolvate thereof.

In a yet further embodiment, the invention provides a compound offormula (I) which comprises1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneor a tautomeric or a stereochemically isomeric form, a pharmaceuticallyacceptable salt or a solvate thereof.

In a yet further embodiment, the invention provides a compound offormula (I) which comprises1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onehydrochloride salt or a tautomeric or a stereochemically isomeric form,or a solvate thereof.

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onedihydrochloride (E2).

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onelactate salt or a tautomeric or a stereochemically isomeric form, or asolvate thereof.

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate salt or a tautomeric or a stereochemically isomeric form,or a solvate thereof.

In a yet further embodiment, the compound is selected from Examples38-42.

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate (Form A) (E39).

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate (Form B) (E40).

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate (Form C) (E43).

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate salt or a tautomeric or a stereochemically isomeric form, or asolvate thereof.

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate (Form F) (E41).

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate salt or a tautomeric or a stereochemically isomeric form, or asolvate thereof.

In a yet further embodiment, the invention provides1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate (Form B) (E42).

In a further embodiment, the compound selected is other than Example 35,or a tautomeric or a stereochemically isomeric form, a pharmaceuticallyacceptable salt or a solvate thereof.

In a further embodiment, the compound selected is other than Example 2,or a tautomeric or a stereochemically isomeric form, a pharmaceuticallyacceptable salt or a solvate thereof.

For the avoidance of doubt, it is to be understood that each general andspecific preference, embodiment and example for one substituent may becombined with each general and specific preference, embodiment andexample for one or more, particularly, all other substituents as definedherein and that all such embodiments are embraced by this application.

Salts, Solvates, Tautomers, Isomers, N-Oxides, Esters, Prodrugs andIsotopes

A reference to a compound of the formula (I) and sub-groups thereof alsoincludes ionic forms, salts, solvates, isomers (including geometric andstereochemical isomers), tautomers, N-oxides, esters, prodrugs, isotopesand protected forms thereof, for example, as discussed below;particularly, the salts or tautomers or isomers or N-oxides or solvatesthereof; and more particularly, the salts or tautomers or N-oxides orsolvates thereof, even more particularly the salts or tautomers orsolvates thereof.

Salts

Many compounds of the formula (I) can exist in the form of salts, forexample acid addition salts or, in certain cases salts of organic andinorganic bases such as carboxylate, sulfonate and phosphate salts. Allsuch salts are within the scope of this invention, and references tocompounds of the formula (I) include the salt forms of the compounds.

The salts of the present invention can be synthesized from the parentcompound that contains a basic or acidic moiety by conventional chemicalmethods such as methods described in Pharmaceutical Salts: Properties,Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth(Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002.Generally, such salts can be prepared by reacting the free acid or baseforms of these compounds with the appropriate base or acid in water orin an organic solvent, or in a mixture of the two; generally, nonaqueousmedia such as ether, ethyl acetate, ethanol, isopropanol, oracetonitrile are used.

Acid addition salts (mono- or di-salts) may be formed with a widevariety of acids, both inorganic and organic. Examples of acid additionsalts include mono- or di-salts formed with an acid selected from thegroup consisting of acetic, 2,2-dichloroacetic, adipic, alginic,ascorbic (e.g. L-ascorbic), L-aspartic, benzenesulfonic, benzoic,4-acetamidobenzoic, butanoic, (+) camphoric, camphor-sulfonic,(+)-(1S)-camphor-10-sulfonic, capric, caproic, caprylic, cinnamic,citric, cyclamic, dodecylsulfuric, ethane-1,2-disulfonic,ethanesulfonic, 2-hydroxyethanesulfonic, formic, fumaric, galactaric,gentisic, glucoheptonic, D-gluconic, glucuronic (e.g. D-glucuronic),glutamic (e.g. L-glutamic), α-oxoglutaric, glycolic, hippuric,hydrohalic acids (e.g. hydrobromic, hydrochloric, hydriodic),isethionic, lactic (e.g. (+)-L-lactic, (±)-DL-lactic), lactobionic,maleic, malic, (−)-L-malic, malonic, (±)-DL-mandelic, methanesulfonic,naphthalene-2-sulfonic, naphthalene-1,5-disulfonic,1-hydroxy-2-naphthoic, nicotinic, nitric, oleic, orotic, oxalic,palmitic, pamoic, phosphoric, propionic, pyruvic, L-pyroglutamic,salicylic, 4-amino-salicylic, sebacic, stearic, succinic, sulfuric,tannic, (+)-L-tartaric, thiocyanic, p-toluenesulfonic, undecylenic andvaleric acids, as well as acylated amino acids and cation exchangeresins.

One particular group of salts consists of salts formed from acetic,hydrochloric, hydriodic, phosphoric, nitric, sulfuric, citric, lactic,succinic, maleic, malic, isethionic, fumaric, benzenesulfonic,toluenesulfonic, methanesulfonic (mesylate), ethanesulfonic,naphthalenesulfonic, valeric, acetic, propanoic, butanoic, malonic,glucuronic and lactobionic acids. One particular, sub group of saltsconsists of salts formed from hydrochloric, lactic (e.g. (+)-L-lactic,(−)-D-lactic or (±)-DL-lactic), sulfuric and methanesulfonic (mesylate)acids. One particular, further sub group of salts consists of saltsformed from lactic (e.g. (+)-L-lactic, (−)-D-lactic or (±)-DL-lactic),sulfuric and methanesulfonic (mesylate) acids. One particular, furthersub group of salts consists of salts formed from lactic (e.g.(+)-L-lactic, (−)-D-lactic or (±)-DL-lactic) and sulfuric acids. Oneparticular salt is the hydrochloride salt. One further particular saltis the lactate salt (such as the compound of Examples 39, 40 and 43).One further particular salt is the sulfate salt (such as the compound ofExample 41). One further particular salt is the mesylate salt (such asthe compound of Example 42). One particular salt is the lactate salt(such as the compound of Examples 39, 40 and 43, in particular thecompound of Example 43), e.g. the L-(+)-lactate salt.

If the compound is anionic, or has a functional group which may beanionic (e.g., —COOH may be —COO⁻), then a salt may be formed with anorganic or inorganic base, generating a suitable cation. Examples ofsuitable inorganic cations include, but are not limited to, alkali metalions such as Li⁺, Na⁺ and K⁺, alkaline earth metal cations such as Ca²⁺and Mg²⁺, and other cations such as Al³⁺ or Zn⁺. Examples of suitableorganic cations include, but are not limited to, ammonium ion (i.e.,NH₄+) and substituted ammonium ions (e.g., NH₃R⁺, NH₂R₂ ⁺, NHR₃ ⁺, NR₄⁺). Examples of some suitable substituted ammonium ions are thosederived from: methylamine, ethylamine, diethylamine, propylamine,dicyclohexylamine, triethylamine, butylamine, ethylenediamine,ethanolamine, diethanolamine, piperazine, benzylamine,phenylbenzylamine, choline, meglumine, and tromethamine, as well asamino acids, such as lysine and arginine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

Where the compounds of the formula (I) contain an amine function, thesemay form quaternary ammonium salts, for example by reaction with analkylating agent according to methods well known to the skilled person.Such quaternary ammonium compounds are within the scope of formula (I).

The compounds of the invention may exist as mono-, di- or tri-salts, inparticular mono- or di-salts, depending upon the pKa of the acid fromwhich the salt is formed.

The salt forms of the compounds of the invention are typicallypharmaceutically acceptable salts, and examples of pharmaceuticallyacceptable salts are discussed in Berge et al., 1977, “PharmaceuticallyAcceptable Salts,” J. Pharm. Sci., Vol. 66, pp. 1-19. However, saltsthat are not pharmaceutically acceptable may also be prepared asintermediate forms which may then be converted into pharmaceuticallyacceptable salts. Such non-pharmaceutically acceptable salt forms, whichmay be useful, for example, in the purification or separation of thecompounds of the invention, also form part of the invention.

In one embodiment of the invention, there is provided a pharmaceuticalcomposition comprising a solution (e.g. an aqueous solution) containinga compound of the formula (I) and sub-groups and examples thereof asdescribed herein in the form of a salt in a concentration of greaterthan 10 mg/mL, typically greater than 15 mg/mL and particularly greaterthan 20 mg/mL.

N-Oxides

Compounds of the formula (I) containing an amine function may also formN-oxides. A reference herein to a compound of the formula (I) thatcontains an amine function also includes the N-oxide.

Where a compound contains several amine functions, one or more than onenitrogen atom may be oxidised to form an N-oxide. Particular examples ofN-oxides are the N-oxides of a tertiary amine or a nitrogen atom of anitrogen-containing heterocycle.

N-Oxides can be formed by treatment of the corresponding amine with anoxidizing agent such as hydrogen peroxide or a per-acid (e.g. aperoxycarboxylic acid), see for example Advanced Organic Chemistry, byJerry March, 4th Edition, Wiley Interscience, pages. More particularly,N-oxides can be made by the procedure of L. W. Deady (Syn. Comm. 1977,7, 509-514) in which the amine compound is reacted withm-chloroperoxybenzoic acid (MCPBA), for example, in an inert solventsuch as dichloromethane.

Geometric Isomers and Tautomers

Compounds of the formula (I) may exist in a number of differentgeometric isomeric, and tautomeric forms and references to compounds ofthe formula (I) include all such forms. For the avoidance of doubt,where a compound can exist in one of several geometric isomeric ortautomeric forms and only one is specifically described or shown, allothers are nevertheless embraced by formula (I).

For example, in compounds of the formula (I), the phenyl ring ofcompounds when X represents NH and U represents carbon can exist in atautomeric form as illustrated below. For simplicity, the generalformula (I) illustrates one form 1 but the formula is to be taken asembracing both tautomeric forms (1 and 2).

Other examples of tautomeric forms include, for example, keto-, enol-,and enolate-forms, as in, for example, the following tautomeric pairs:keto/enol (illustrated below), imine/enamine, amide/imino alcohol,amidine/enediamines, nitroso/oxime, thioketone/enethiol, andnitro/aci-nitro.

Stereoisomers

Unless otherwise mentioned or indicated, the chemical designation ofcompounds denotes the mixture of all possible stereochemically isomericforms.

Stereocentres are illustrated in the usual fashion, using ‘hashed’ or‘wedged’ lines. e.g.

Where a compound is described as a mixture of twodiastereoisomers/epimers, the configuration of the stereocentre is notspecified and is represented by straight lines.

Unless otherwise mentioned or indicated, where compounds of the formula(I) contain one or more chiral centres, and can exist in the form of twoor more optical isomers, references to compounds of the formula (I)include all optical isomeric forms thereof (e.g. enantiomers, epimersand diastereoisomers), either as individual optical isomers, or mixtures(e.g. racemic mixtures) or two or more optical isomers, unless thecontext requires otherwise.

The optical isomers may be characterised and identified by their opticalactivity (i.e. as + and − isomers, or d and l isomers) or they may becharacterised in terms of their absolute stereochemistry using the “Rand S” nomenclature developed by Cahn, Ingold and Prelog, see AdvancedOrganic Chemistry by Jerry March, 4th Edition, John Wley & Sons, NewYork, 1992, pages 109-114, and see also Cahn, Ingold & Prelog, Angew.Chem. Int. Ed. Engl., 1966, 5, 385-415.

Optical isomers can be separated by a number of techniques includingchiral chromatography (chromatography on a chiral support) and suchtechniques are well known to the person skilled in the art.

As an alternative to chiral chromatography, optical isomers can beseparated by forming diastereoisomeric salts with chiral acids such as(+)-tartaric acid, (−)-pyroglutamic acid, (−)-di-toluoyl-L-tartaricacid, (+)-mandelic acid, (−)-malic acid, and (−)-camphorsulfonic acid,separating the diastereoisomers by preferential crystallisation, andthen dissociating the salts to give the individual enantiomer of thefree base.

Additionally enantiomeric separation can be achieved by covalentlylinking a enantiomerically pure chiral auxiliary onto the compound andthen performing diastereisomer separation using conventional methodssuch as chromatography. This is then followed by cleavage of theaforementioned covalent linkage to generate the appropriateenantiomerically pure product.

Where compounds of the formula (I) exist as two or more optical isomericforms, one enantiomer in a pair of enantiomers may exhibit advantagesover the other enantiomer, for example, in terms of biological activity.Thus, in certain circumstances, it may be desirable to use as atherapeutic agent only one of a pair of enantiomers, or only one of aplurality of diastereoisomers. Accordingly, the invention providescompositions containing a compound of the formula (I) having one or morechiral centres, wherein at least 55% (e.g. at least 60%, 65%, 70%, 75%,80%, 85%, 90% or 95%) of the compound of the formula (I) is present as asingle optical isomer (e.g. enantiomer or diastereoisomer). In onegeneral embodiment, 99% or more (e.g. substantially all) of the totalamount of the compound of the formula (I) may be present as a singleoptical isomer (e.g. enantiomer or diastereoisomer).

Compounds encompassing double bonds can have an E (entgegen) or Z(zusammen) stereochemistry at said double bond. Substituents on bivalentcyclic or (partially) saturated radicals may have either the cis- ortrans-configuration. The terms cis and trans when used herein are inaccordance with Chemical Abstracts nomenclature (J. Org. Chem. 1970, 35(9), 2849-2867), and refer to the position of the substituents on a ringmoiety.

Of special interest are those compounds of formula (I) which arestereochemically pure. When a compound of formula (I) is for instancespecified as R, this means that the compound is substantially free ofthe S isomer. If a compound of formula (I) is for instance specified asE, this means that the compound is substantially free of the Z isomer.The terms cis, trans, R, S, E and Z are well known to a person skilledin the art.

Isotopic Variations

The present invention includes all pharmaceutically acceptableisotopically-labeled compounds of the invention, i.e. compounds offormula (I), wherein one or more atoms are replaced by atoms having thesame atomic number, but an atomic mass or mass number different from theatomic mass or mass number usually found in nature.

Examples of isotopes suitable for inclusion in the compounds of theinvention comprise isotopes of hydrogen, such as ²H (D) and ³H (T),carbon, such as ¹¹C, ¹³C and ¹⁴C, chlorine, such as ³⁶Cl, fluorine, suchas ¹⁸F, iodine, such as ¹²³I, ¹²⁵I and ¹³¹I nitrogen, such as ¹³N and¹⁵N, oxygen, such as ¹⁵O, ¹⁷O and ¹⁸O, phosphorus, such as ³²P, andsulfur, such as ³⁵S.

Certain isotopically-labelled compounds of formula (I), for example,those incorporating a radioactive isotope, are useful in drug and/orsubstrate tissue distribution studies. The compounds of formula (I) canalso have valuable diagnostic properties in that they can be used fordetecting or identifying the formation of a complex between a labelledcompound and other molecules, peptides, proteins, enzymes or receptors.The detecting or identifying methods can use compounds that are labelledwith labelling agents such as radioisotopes, enzymes, fluorescentsubstances, luminous substances (for example, luminol, luminolderivatives, luciferin, aequorin and luciferase), etc. The radioactiveisotopes tritium, i.e. ³H (T), and carbon-14, i.e. ¹⁴C, are particularlyuseful for this purpose in view of their ease of incorporation and readymeans of detection.

Substitution with heavier isotopes such as deuterium, i.e. ²H (D), mayafford certain therapeutic advantages resulting from greater metabolicstability, for example, increased in vivo half-life or reduced dosagerequirements, and hence may be used in some circumstances.

Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O and¹³N, can be useful in Positron Emission Topography (PET) studies forexamining target occupancy.

Isotopically-labeled compounds of formula (I) can generally be preparedby conventional techniques known to those skilled in the art or byprocesses analogous to those described in the accompanying Examples andPreparations using an appropriate isotopically-labeled reagents in placeof the non-labeled reagent previously employed.

Esters

Esters such as carboxylic acid esters, acyloxy esters and phosphateesters of the compounds of formula (I) bearing a carboxylic acid groupor a hydroxyl group are also embraced by Formula (I). Examples of estersare compounds containing the group —C(═O)OR, wherein R is an estersubstituent, for example, a C₁₋₇ alkyl group, a C₃₋₁₂ heterocyclylgroup, or a C₅₋₁₂ aryl group, particularly a O₁₋₆ alkyl group.Particular examples of ester groups include, but are not limited to—C(═O)OCH₃, —C(═O)OCH₂CH₃, —C(═O)OC(CH₃)₃, and —C(═O)OPh. Examples ofacyloxy (reverse ester) groups are represented by —OC(═O)R, wherein R isan acyloxy substituent, for example, a c₁₋₆ alkyl group, a C₃₋₁₂heterocyclyl group, or a C₅₋₁₂ aryl group, particularly a C₁₋₆ alkylgroup. Particular examples of acyloxy groups include, but are notlimited to, —OC(═O)CH₃ (acetoxy), —OC(═O)CH₂CH₃, —OC(═O)C(CH₃)₃,—OC(═O)Ph, and —OC(═O)CH₂Ph. Examples of phosphate esters are thosederived from phosphoric acid.

In one embodiment of the invention, formula (I) includes within itsscope esters of compounds of the formula (I) bearing a carboxylic acidgroup or a hydroxyl group. In another embodiment of the invention,formula (I) does not include within its scope esters of compounds of theformula (I) bearing a carboxylic acid group or a hydroxyl group.

Solvates and Crystalline Forms

Also encompassed by formula (I) are any polymorphic forms of thecompounds, and solvates such as hydrates, alcoholates and the like.

The compounds of the invention may form solvates, for example with water(i.e., hydrates) or common organic solvents. As used herein, the term“solvate” means a physical association of the compounds of the presentinvention with one or more solvent molecules. This physical associationinvolves varying degrees of ionic and covalent bonding, includinghydrogen bonding. In certain instances the solvate will be capable ofisolation, for example when one or more solvent molecules areincorporated in the crystal lattice of the crystalline solid. The term“solvate” is intended to encompass both solution-phase and isolatablesolvates. Non-limiting examples of suitable solvates include compoundsof the invention in combination with water, isopropanol, ethanol,methanol, DMSO, ethyl acetate, acetic acid or ethanolamine and the like.The compounds of the invention may exert their biological effects whilstthey are in solution.

Solvates are well known in pharmaceutical chemistry. They can beimportant to the processes for the preparation of a substance (e.g. inrelation to their purification, the storage of the substance (e.g. itsstability) and the ease of handling of the substance and are oftenformed as part of the isolation or purification stages of a chemicalsynthesis. A person skilled in the art can determine by means ofstandard and long used techniques whether a hydrate or other solvate hasformed by the isolation conditions or purification conditions used toprepare a given compound. Examples of such techniques includethermogravimetric analysis (TGA), differential scanning calorimetry(DSC), X-ray crystallography (e.g. single crystal X-ray crystallographyor X-ray powder diffraction) and Solid State NMR (SS-NMR, also known asMagic Angle Spinning NMR or MAS-NMR). Such techniques are as much a partof the standard analytical toolkit of the skilled chemist as NMR, IR,HPLC and MS.

Alternatively the skilled person can deliberately form a solvate usingcrystallisation conditions that include an amount of the solventrequired for the particular solvate. Thereafter the standard methodsdescribed above, can be used to establish whether solvates had formed.

In one embodiment the salt of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onehas <10% solvates present (such as no more than any one of the followingamounts 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.1, 0.05 or 0.01%), e.g.hydrates, alcoholates, isopropylacetate, methyl acetate or alkanes, suchas heptanes.

In one embodiment the salt of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneis anhydrous. In a further embodiment, the anhydrous salt of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onecontains no more than 5% (such as no more than any one of the followingamounts 4, 3, 2, 1, 0.5, 0.1, 0.05 or 0.01%) by weight of water.

In one embodiment the salt of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onecontains a single crystalline form and no more than 5% (such as no morethan any one of the following amounts 4, 3, 2, 1, 0.5, 0.1, 0.05 or0.01%) by weight of other crystalline forms.

In one embodiment the salt of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneis crystalline.

In one embodiment the salt of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneis amorphous.

Furthermore, the compounds of the present invention may have one or morepolymorph or amorphous crystalline forms and as such are intended to beincluded in the scope of the invention.

References herein to “polymorph” refer to the existence of more than onecrystal structure of a compound of formula (I). The ability of achemical compound to crystallize in more than one crystal modificationcan have an effect upon the properties of said compound, such asphysicochemical properties, shelf life, solubility, formulationproperties, toxicity, bioavailability, hygroscopicity and processingproperties. In addition, the therapeutic action of a pharmaceuticalcompound can be affected by the polymorphism of the drug molecule.

In one embodiment, the compound of formula (I) comprises a polymorphicform of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneor salt thereof.

In a further embodiment, the compound of formula (I) comprises apolymorphic form of a salt of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one.

In a further embodiment, the compound of formula (I) comprises the FormA polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate. This compound may be prepared as defined herein inExample 39.

In a yet further embodiment,1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by the ¹H NMR spectrum depicted in FIG.1.

A compound's X-ray powder pattern is characterised by the diffractionangle (2θ) and interplanar spacing (d) parameters of an X-raydiffraction spectrum. These are related by Bragg's equation, nλ=2d Sinθ, (where n=1; λ=wavelength of the cathode used; d=interplanar spacing;and θ=diffraction angle). Herein, interplanar spacings, diffractionangle and overall pattern are important for identification of crystal inthe X-ray powder diffraction, due to the characteristics of the data.The relative intensity should not be strictly interpreted since it maybe varied depending on the direction of crystal growth, particle sizesand measurement conditions. In addition, the diffraction angles usuallymean ones which coincide in the range of 2θ±0.2°. The peaks mean mainpeaks and include peaks not larger than medium at diffraction anglesother than those stated above.

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at6.5±0.5°, 7.1±0.5°, 7.9±0.5°, 9.3±0.5°, 10.2±0.5°, 11.0±0.5°, 11.6±0.5°,13.3±0.5°, 14.4±0.5°, 15.0±0.5°, 16.7±0.5°, 18.0±0.5°, 18.4±0.5°,20.0±0.5°, 21.0±0.5°, 23.4±0.5°, 25.2±0.5° and 26.1±0.5° (2θ, 1d.p).

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at6.5±0.2°, 7.1±0.2°, 7.9±0.2°, 9.3±0.2°, 10.2±0.2°, 11.0±0.2°, 11.6±0.2°,13.3±0.2°, 14.4±0.2°, 15.0±0.2°, 16.7±0.2°, 18.0±0.2°, 18.4±0.2°,20.0±0.2°, 21.0±0.2°, 23.4±0.2°, 25.2±0.2° and 26.1±0.2° (2θ, 1d.p).

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at6.5±0.1°, 7.1±0.1°, 7.9±0.1°, 9.3±0.1°, 10.2±0.1°, 11.0±0.1°, 11.6±0.1°,13.3±0.1°, 14.4±0.1°, 15.0±0.1°, 16.7±0.1°, 18.0±0.1°, 18.4±0.1°,20.0±0.1°, 21.0±0.1°, 23.4±0.1°, 25.2±0.1° and 26.1±0.1° (2θ, 1d.p).

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at 6.5°,7.1°, 7.9°, 9.3°, 10.2°, 11.0°, 11.6°, 13.3°, 14.4°, 15.0°, 16.7°,18.0°, 18.4°, 20.0°, 21.0°, 23.4°, 25.2° and 26.1° (2θ, 1d.p).

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern substantially as shownin FIG. 2.

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having peaks at the same diffractionangles (2θ) of the XRPD pattern shown in FIG. 2 and optionally whereinthe peaks have the same relative intensity as the peaks shown in FIG. 2.

It will be appreciated by the skilled person that references herein to“intensity” of peaks with respect to XRPD refer to relative intensitieswhich have taken into account normalisation of background noise andother such parameters.

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having major peaks at diffractionangles (2θ) and intensities as those shown in the XRPD pattern in FIG.2.

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of13.59±0.5 Å, 12.44±0.5 Å, 11.19±0.5 Å, 9.50±0.5 Å, 8.67±0.5 Å, 8.04±0.5Å, 7.62±0.5 Å, 6.65±0.5 Å, 6.15±0.5 Å, 5.90±0.5 Å, 5.31±0.5 Å, 4.93±0.5Å, 4.82±0.5 Å, 4.44±0.5 Å, 4.23±0.5 Å, 3.80±0.5 Å, 3.53±0.5 Å and3.41±0.5 Å (d, 2d.p.).

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of13.59±0.2 Å, 12.44±0.2 Å, 11.19±0.2 Å, 9.50±0.2 Å, 8.67±0.2 Å, 8.04±0.2Å, 7.62±0.2 Å, 6.65±0.2 Å, 6.15±0.2 Å, 5.90±0.2 Å, 5.31±0.2 Å, 4.93±0.2Å, 4.82±0.2 Å, 4.44±0.2 Å, 4.23±0.2 Å, 3.80±0.2 Å, 3.53±0.2 Å and3.41±0.2 Å (d, 2d.p.).

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of13.59±0.1 Å, 12.44±0.1 Å, 11.19±0.1 Å, 9.50±0.1 Å, 8.67±0.1 Å, 8.04±0.1Å, 7.62±0.1 Å, 6.65±0.1 Å, 6.15±0.1 Å, 5.90±0.1 Å, 5.31±0.1 Å, 4.93±0.1Å, 4.82±0.1 Å, 4.44±0.1 Å, 4.23±0.1 Å, 3.80±0.1 Å, 3.53±0.1 Å and3.41±0.1 Å (d, 2d.p.).

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of13.59 Å, 12.44 Å, 11.19 Å, 9.50 Å, 8.67 Å, 8.04 Å, 7.62 Å, 6.65 Å, 6.15Å, 5.90 Å, 5.31 Å, 4.93 Å, 4.82 Å, 4.44 Å, 4.23 Å, 3.80 Å, 3.53 Å and3.41 Å (d, 2d.p.).

In a further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by DSC peak temperatures of 78.69°C.±0.5° C. and/or 113.91° C.±0.5° C. (such as 78.69° C.±0.2° C. and/or113.91° C.±0.2° C., in particular 78.69° C.±0.1° C. and/or 113.91°C.±0.1° C., more particularly 78.69° C. and/or 113.91° C.).

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by DSC onset temperatures of 72.3°C.±0.5° C. (endotherm, broad) and/or 102° C.±0.5° C. (endotherm, broad)(such as 72.3° C.±0.2° C. and/or 102° C.±0.2° C., in particular 72.3°C.±0.1° C. and/or 102° C.±0.1° C., more particularly 72.3° C. and/or102° C.).

In a yet further embodiment, the Form A polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by a DSC thermogram as depicted in FIG.3.

In a further embodiment, the compound of formula (I) comprises the FormB polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate. This compound may be prepared as defined herein inExample 40.

In a yet further embodiment,1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by the ¹H NMR spectrum depicted in FIG.4.

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at6.6±0.5°, 9.4±0.5°, 11.0±0.5°, 13.2±0.5°, 14.3±0.5°, 15.8±0.5°,17.4±0.5°, 18.4±0.5°, 19.1±0.5°, 20.9±0.5°, 21.8±0.5°, 23.1±0.5°,24.9±0.5°, 26.7±0.5° and 27.8±0.5° (2θ, 1d.p).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at6.6±0.2°, 9.4±0.2°, 11.0±0.2°, 13.2±0.2°, 14.3±0.2°, 15.8±0.2°,17.4±0.2°, 18.4±0.2°, 19.1±0.2°, 20.9±0.2°, 21.8±0.2°, 23.1±0.2°,24.9±0.2°, 26.7±0.2° and 27.8±0.2° (2θ, 1d.p).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an

XRPD pattern having peaks at 6.6±0.1°, 9.4±0.1°, 11.0±0.1°, 13.2±0.1°,14.3±0.1°, 15.8±0.1°, 17.4±0.1°, 18.4±0.1°, 19.1±0.1°, 20.9±0.1°,21.8±0.1°, 23.1±0.1°, 24.9±0.1°, 26.7±0.1° and 27.8±0.1° (2θ, 1d.p).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at 6.6°,9.4°, 11.0°, 13.2°, 14.3°, 15.8°, 17.4°, 18.4°, 19.1°, 20.9°, 21.8°,23.1°, 24.9°, 26.7° and 27.8° (2θ, 1d.p).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern substantially as shownin FIG. 5.

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having peaks at the same diffractionangles (2θ) of the XRPD pattern shown in FIG. 5 and optionally whereinthe peaks have the same relative intensity as the peaks shown in FIG. 5.

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having major peaks at diffractionangles (2θ) and intensities as those shown in the XRPD pattern in FIG.5.

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of13.39±0.5 Å, 9.40±0.5 Å, 8.04±0.5 Å, 6.70±0.5 Å, 6.19±0.5 Å, 5.61±0.5 Å,5.09±0.5 Å, 4.82±0.5 Å, 4.64±0.5 Å, 4.25±0.5 Å, 4.07±0.5 Å, 3.85±0.5 Å,3.57±0.5 Å, 3.34±0.5 Å and 3.21±0.5 Å (d, 2d.p.).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of13.39±0.2 Å, 9.40±0.2 Å, 8.04±0.2 Å, 6.70±0.2 Å, 6.19±0.2 Å, 5.61±0.2 Å,5.09±0.2 Å, 4.82±0.2 Å, 4.64±0.2 Å, 4.25±0.2 Å, 4.07±0.2 Å, 3.85±0.2 Å,3.57±0.2 Å, 3.34±0.2 Å and 3.21±0.2 Å (d, 2d.p.).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of13.39±0.1 Å, 9.40±0.1 Å, 8.04±0.1 Å, 6.70±0.1 Å, 6.19±0.1 Å, 5.61±0.1 Å,5.09±0.1 Å, 4.82±0.1 Å, 4.64±0.1 Å, 4.25±0.1 Å, 4.07±0.1 Å, 3.85±0.1 Å,3.57±0.1 Å, 3.34±0.1 Å and 3.21±0.1 Å (d, 2d.p.).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of13.39 Å, 9.40 Å, 8.04 Å, 6.70 Å, 6.19 Å, 5.61 Å, 5.09 Å, 4.82 Å, 4.64 Å,4.25 Å, 4.07 Å, 3.85 Å, 3.57 Å, 3.34 Å and 3.21 Å (d, 2d.p.).

In a further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by DSC peak temperatures of 85.25°C.±0.5° C. and/or 106.72° C.±0.5° C. (such as 85.25° C.±0.2° C. and/or106.72° C.±0.2° C., in particular 85.25° C.±0.1° C. and/or 106.72°C.±0.1° C., more particularly 85.25° C. and/or 106.72° C.).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by DSC onset temperatures of 68° C.±0.5°C. (large endotherm, broad) and/or 102° C.±0.5° C. (very smallendotherm, broad) (such as 68° C.±0.2° C. and/or 102° C.±0.2° C., inparticular 68° C.±0.1° C. and/or 102° C.±0.1° C., more particularly 68°C. and/or 102° C.).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by a DSC thermogram as depicted in FIG.6.

In a further embodiment, the compound of formula (I) comprises the FormF polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate. This compound may be prepared as defined herein in Example 41.

In a yet further embodiment,1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by the ¹H NMR spectrum depicted in FIG. 7.

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by an XRPD pattern having peaks at 8.5±0.5°,13.5±0.5°, 13.9±0.5°, 14.3±0.5°, 16.2±0.5°, 17.3±0.5°, 20.1±0.5°,21.3±0.5°, 23.3±0.5°, 24.4±0.5° and 27.9±0.5° (2θ, 1d.p).

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by an XRPD pattern having peaks at 8.5±0.2°,13.5±0.2°, 13.9±0.2°, 14.3±0.2°, 16.2±0.2°, 17.3±0.2°, 20.1±0.2°,21.3±0.2°, 23.3±0.2°, 24.4±0.2° and 27.9±0.2° (2θ, 1d.p).

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by an XRPD pattern having peaks at 8.5±0.1°,13.5±0.1°, 13.9±0.1°, 14.3±0.1°, 16.2±0.1°, 17.3±0.1°, 20.1±0.1°,21.3±0.1°, 23.3±0.1°, 24.4±0.1° and 27.9±0.1° (2θ, 1d.p).

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by an XRPD pattern having peaks at 8.5°, 13.5°,13.9°, 14.3°, 16.2°, 17.3°, 20.1°, 21.3°, 23.3°, 24.4° and 27.9° (2θ,1d.p).

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by an XRPD pattern substantially as shown inFIG. 8.

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by having peaks at the same diffraction angles(2θ) of the XRPD pattern shown in FIG. 8 and optionally wherein thepeaks have the same relative intensity as the peaks shown in FIG. 8.

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by having major peaks at diffraction angles(2θ) and intensities as those shown in the XRPD pattern in FIG. 8.

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by interplanar spacing (d) values of 10.40±0.5Å, 6.56±0.5 Å, 6.37±0.5 Å, 6.19±0.5 Å, 5.47±0.5 Å, 5.12±0.5 Å, 4.42±0.5Å, 4.17±0.5 Å, 3.82±0.5 Å, 3.65±0.5 Å and 3.20±0.5 Å (d, 2d.p.).

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by interplanar spacing (d) values of 10.40±0.2Å, 6.56±0.2 Å, 6.37±0.2 Å, 6.19±0.2 Å, 5.47±0.2 Å, 5.12±0.2 Å, 4.42±0.2Å, 4.17±0.2 Å, 3.82±0.2 Å, 3.65±0.2 Å and 3.20±0.2 Å (d, 2d.p.).

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by interplanar spacing (d) values of 10.40±0.1Å, 6.56±0.1 Å, 6.37±0.1 Å, 6.19±0.1 Å, 5.47±0.1 Å, 5.12±0.1 Å, 4.42±0.1Å, 4.17±0.1 Å, 3.82±0.1 Å, 3.65±0.1 Å and 3.20±0.1 Å (d, 2d.p.).

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by interplanar spacing (d) values of 10.40 Å,6.56 Å, 6.37 Å, 6.19 Å, 5.47 Å, 5.12 Å, 4.42 Å, 4.17 Å, 3.82 Å, 3.65 Åand 3.20 Å (d, 2d.p.).

In a further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by DSC peak temperatures of 80.31° C.±0.5° C.and/or 149.07° C.±0.5° C. (such as 80.31° C.±0.2° C. and/or 149.07°C.±0.2° C., in particular 80.31° C.±0.1° C. and/or 149.07° C.±0.1° C.,more particularly 80.31° C. and/or 149.07° C.).

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by DSC onset temperatures of 51.2° C.±0.5° C.(endotherm, broad) and/or 136° C.±0.5° C. (endotherm, broad) (such as51.2° C.±0.2° C. and/or 136° C.±0.2° C., in particular 51.2° C.±0.1° C.and/or 136° C.±0.1° C., more particularly 51.2° C. and/or 136° C.).

In a yet further embodiment, the Form F polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate is characterised by a DSC thermogram as depicted in FIG. 9.

In a further embodiment, the compound of formula (I) comprises the FormB polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate. This compound may be prepared as defined herein in Example 42.

In a yet further embodiment,1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by the ¹H NMR spectrum depicted in FIG. 10.

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by an XRPD pattern having peaks at 6.6±0.5°,8.0±0.5°, 11.8±0.5°, 13.2±0.5°, 14.3±0.5°, 15.0±0.5°, 15.6±0.5°,17.1±0.5°, 17.4±0.5°, 17.7±0.5°, 19.2±0.5°, 20.3±0.5°, 21.2±0.5°,22.3±0.5°, 23.0±0.5°, 24.0±0.5°, 25.8±0.5°, 26.8±0.5° and 28.9±0.5° (2θ,1d.p).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by an XRPD pattern having peaks at 6.6±0.2°,8.0±0.2°, 11.8±0.2°, 13.2±0.2°, 14.3±0.2°, 15.0±0.2°, 15.6±0.2°,17.1±0.2°, 17.4±0.2°, 17.7±0.2°, 19.2±0.2°, 20.3±0.2°, 21.2±0.2°,22.3±0.2°, 23.0±0.2°, 24.0±0.2°, 25.8±0.2°, 26.8±0.2° and 28.9±0.2° (2θ,1d.p).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by an

XRPD pattern having peaks at 6.6±0.1°, 8.0±0.1°, 11.8±0.1°, 13.2±0.1°,14.3±0.1°, 15.0±0.1°, 15.6±0.1°, 17.1±0.1°, 17.4±0.1°, 17.7±0.1°,19.2±0.1°, 20.3±0.1°, 21.2±0.1°, 22.3±0.1°, 23.0±0.1°, 24.0±0.1°,25.8±0.1°, 26.8±0.1° and 28.9±0.1° (2θ, 1d.p).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by an XRPD pattern having peaks at 6.6°, 8.0°,11.8°, 13.2°, 14.3°, 15.0°, 15.6°, 17.1°, 17.4°, 17.7°, 19.2°, 20.3°,21.2°, 22.3°, 23.0°, 24.0°, 25.8°, 26.8° and 28.9° (2θ, 1d.p).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by an XRPD pattern substantially as shown inFIG. 11.

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by having peaks at the same diffraction angles(2θ) of the XRPD pattern shown in FIG. 11 and optionally wherein thepeaks have the same relative intensity as the peaks shown in FIG. 11.

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by having major peaks at diffraction angles(2θ) and intensities as those shown in the XRPD pattern in FIG. 11.

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by interplanar spacing (d) values of 13.39±0.5Å, 11.05±0.5 Å, 7.50±0.5 Å, 6.70±0.5 Å, 6.19±0.5 Å, 5.90±0.5 Å, 5.68±0.5Å, 5.18±0.5 Å, 5.09±0.5 Å, 5.01±0.5 Å, 4.62±0.5 Å, 4.37±0.5 Å, 4.19±0.5Å, 3.98±0.5 Å, 3.86±0.5 Å, 3.71±0.5 Å, 3.45±0.5 Å, 3.32±0.5 Å and3.09±0.5 Å (d, 2d.p.).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by interplanar spacing (d) values of 13.39±0.2Å, 11.05±0.2 Å, 7.50±0.2 Å, 6.70±0.2 Å, 6.19±0.2 Å, 5.90±0.2 Å, 5.68±0.2Å, 5.18±0.2 Å, 5.09±0.2 Å, 5.01±0.2 Å, 4.62±0.2 Å, 4.37±0.2 Å, 4.19±0.2Å, 3.98±0.2 Å, 3.86±0.2 Å, 3.71±0.2 Å, 3.45±0.2 Å, 3.32±0.2 Å and3.09±0.2 Å (d, 2d.p.).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by interplanar spacing (d) values of 13.39±0.1Å, 11.05±0.1 Å, 7.50±0.1 Å, 6.70±0.1 Å, 6.19±0.1 Å, 5.90±0.1 Å, 5.68±0.1Å, 5.18±0.1 Å, 5.09±0.1 Å, 5.01±0.1 Å, 4.62±0.1 Å, 4.37±0.1 Å, 4.19±0.1Å, 3.98±0.1 Å, 3.86±0.1 Å, 3.71±0.1 Å, 3.45±0.1 Å, 3.32±0.1 Å and3.09±0.1 Å (d, 2d.p.).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by interplanar spacing (d) values of 13.39 Å,11.05 Å, 7.50 Å, 6.70 Å, 6.19 Å, 5.90 Å, 5.68 Å, 5.18 Å, 5.09 Å, 5.01 Å,4.62 Å, 4.37 Å, 4.19 Å, 3.98 Å, 3.86 Å, 3.71 Å, 3.45 Å, 3.32 Å and 3.09Å (d, 2d.p.).

In a further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by DSC peak temperatures of 98.63° C.±0.5° C.and/or 177.11° C.±0.5° C. (such as 98.63° C.±0.2° C. and/or 177.11°C.±0.2° C., in particular 98.63° C.±0.1° C. and/or 177.11° C.±0.1° C.,more particularly 98.63° C. and/or 177.11° C.).

In a further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by DSC onset temperatures of 73.3° C.±0.5° C.(endotherm, broad) and/or 160.8° C.±0.5° C. (endotherm, broad) (such as73.3° C.±0.2° C. and/or 160.8° C.±0.2° C., in particular 73.3° C.±0.1°C. and/or 160.8° C.±0.1° C., more particularly 73.3° C. and/or 160.8°C.).

In a yet further embodiment, the Form B polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate is characterised by a DSC thermogram as depicted in FIG. 12.

In a further embodiment, the compound of formula (I) comprises the FormC polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate. This compound may be prepared as defined herein inExample 43.

In a yet further embodiment,1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by the ¹H NMR spectrum depicted in FIG.15.

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at7.4±0.5°, 7.9±0.5°, 8.3±0.5°, 8.7±0.5°, 9.0±0.5°, 10.4±0.5°, 11.2±0.5°,11.6±0.5°, 12.3±0.5°, 13.1±0.5°, 13.9±0.5°, 14.7±0.5°, 15.8±0.5°,16.5±0.5°, 17.1±0.5°, 17.9±0.5°, 18.4±0.5°, 18.9±0.5°, 19.6±0.5°,20.4±0.5°, 21.0±0.5°, 21.8±0.5°, 22.9±0.5°, 23.3±0.5°, 23.6±0.5°,24.0±0.5°, 24.9±0.5° and 26.4±0.5° (2θ, 1d.p).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an

XRPD pattern having peaks at 7.4±0.2°, 7.9±0.2°, 8.3±0.2°, 8.7±0.2°,9.0±0.2°, 10.4±0.2°, 11.2±0.2°, 11.6±0.2°, 12.3±0.2°, 13.1±0.2°,13.9±0.2°, 14.7±0.2°, 15.8±0.2°, 16.5±0.2°, 17.1±0.2°, 17.9±0.2°,18.4±0.2°, 18.9±0.2°, 19.6±0.2°, 20.4±0.2°, 21.0±0.2°, 21.8±0.2°,22.9±0.2°, 23.3±0.2°, 23.6±0.2°, 24.0±0.2°, 24.9±0.2° and 26.4±0.2° (2θ,1d.p).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at7.4±0.1°, 7.9±0.1°, 8.3±0.1°, 8.7±0.1°, 9.0±0.1°, 10.4±0.1°, 11.2±0.1°,11.6±0.1°, 12.3±0.1°, 13.1±0.1°, 13.9±0.1°, 14.7±0.1°, 15.8±0.1°,16.5±0.1°, 17.1±0.1°, 17.9±0.1°, 18.4±0.1°, 18.9±0.1°, 19.6±0.1°,20.4±0.1°, 21.0±0.1°, 21.8±0.1°, 22.9±0.1°, 23.3±0.1°, 23.6±0.1°,24.0±0.1°, 24.9±0.1° and 26.4±0.1° (2θ, 1d.p).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern having peaks at 7.4°,7.9°, 8.3°, 8.7°, 9.0°, 10.4°, 11.2°, 11.6°, 12.3°, 13.1°, 13.9°, 14.7°,15.8°, 16.5°, 17.1°, 17.9°, 18.4°, 18.9°, 19.6°, 20.4°, 21.0°, 21.8°,22.9°, 23.3°, 23.6°, 24.0°, 24.9° and 26.4° (2θ, 1d.p).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by an XRPD pattern substantially as shownin FIG. 16 labelled as 1.

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having peaks at the same diffractionangles (2θ) of the XRPD pattern shown in FIG. 16 labelled as 1 andoptionally wherein the peaks have the same relative intensity as thepeaks shown in FIG. 16 labelled as 1.

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having major peaks at diffractionangles (2θ) and intensities as those shown in the XRPD pattern in FIG.16 labelled as 1.

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having major peaks as measured by XRPDat 8.7±0.5°, 17.1±0.5°, 17.9±0.5° and 18.9±0.5° (2θ, 1d.p).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having major peaks as measured by XRPDat 8.7±0.2°, 17.1±0.2°, 17.9±0.2° and 18.9±0.2° (2θ, 1d.p).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having major peaks as measured by XRPDat 8.7±0.1°, 17.1±0.1°, 17.9±0.1° and 18.9±0.1° (2θ, 1d.p).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by having major peaks as measured by XRPDat 8.7°, 17.1°, 17.9° and 18.9° (2θ, 1d.p).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of11.94±0.5 Å, 11.19±0.5 Å, 10.65±0.5 Å, 10.16±0.5 Å, 9.82±0.5 Å, 8.50±0.5Å, 7.90±0.5 Å, 7.62±0.5 Å, 7.19±0.5 Å, 6.75±0.5 Å, 6.37±0.5 Å, 6.02±0.5Å, 5.61±0.5 Å, 5.37±0.5 Å, 5.18±0.5 Å, 4.95±0.5 Å, 4.82±0.5 Å, 4.69±0.5Å, 4.53±0.5 Å, 4.35±0.5 Å, 4.23±0.5 Å, 4.07±0.5 Å, 3.88±0.5 Å, 3.82±0.5Å, 3.77±0.5 Å, 3.71±0.5 Å, 3.57±0.5 Å and 3.37±0.5 Å (d, 2d.p.).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of11.94±0.2 Å, 11.19±0.2 Å, 10.65±0.2 Å, 10.16±0.2 Å, 9.82±0.2 Å, 8.50±0.2Å, 7.90±0.2 Å, 7.62±0.2 Å, 7.19±0.2 Å, 6.75±0.2 Å, 6.37±0.2 Å, 6.02±0.2Å, 5.61±0.2 Å, 5.37±0.2 Å, 5.18±0.2 Å, 4.95±0.2 Å, 4.82±0.2 Å, 4.69±0.2Å, 4.53±0.2 Å, 4.35±0.2 Å, 4.23±0.2 Å, 4.07±0.2 Å, 3.88±0.2 Å, 3.82±0.2Å, 3.77±0.2 Å, 3.71±0.2 Å, 3.57±0.2 Å and 3.37±0.2 Å (d, 2d.p.).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of11.94±0.1 Å, 11.19±0.1 Å, 10.65±0.1 Å, 10.16±0.1 Å, 9.82±0.1 Å, 8.50±0.1Å, 7.90±0.1 Å, 7.62±0.1 Å, 7.19±0.1 Å, 6.75±0.1 Å, 6.37±0.1 Å, 6.02±0.1Å, 5.61±0.1 Å, 5.37±0.1 Å, 5.18±0.1 Å, 4.95±0.1 Å, 4.82±0.1 Å, 4.69±0.1Å, 4.53±0.1 Å, 4.35±0.1 Å, 4.23±0.1 Å, 4.07±0.1 Å, 3.88±0.1 Å, 3.82±0.1Å, 3.77±0.1 Å, 3.71±0.1 Å, 3.57±0.1 Å and 3.37±0.1 Å (d, 2d.p.).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by interplanar spacing (d) values of11.94 Å, 11.19 Å, 10.65 Å, 10.16 Å, 9.82 Å, 8.50 Å, 7.90 Å, 7.62 Å, 7.19Å, 6.75 Å, 6.37 Å, 6.02 Å, 5.61 Å, 5.37 Å, 5.18 Å, 4.95 Å, 4.82 Å, 4.69Å, 4.53 Å, 4.35 Å, 4.23 Å, 4.07 Å, 3.88 Å, 3.82 Å, 3.77 Å, 3.71 Å, 3.57Å and 3.37 Å (d, 2d.p.).

In a further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by a DSC peak temperature of 174.37°C.±0.5° C. (such as 174.37° C.±0.2° C., in particular 174.37° C.±0.1°C., more particularly 174.37° C.).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by a DSC onset temperature of 171.6°C.±0.5° C. (endotherm, sharp) (such as 171.6° C.±0.2° C., in particular171.6° C.±0.1° C., more particularly 171.6° C.).

In a yet further embodiment, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate is characterised by a DSC thermogram as depicted in FIG.17 labelled as 1.

In one embodiment, a lactate (e.g. L-(+)-lactate) salt of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onewhich is crystalline and is characterised by one or more (in anycombination) or all of the following parameters:

(a) the ¹H NMR spectrum depicted in FIG. 15; and/or(b) an XRPD pattern having peaks at 7.4±0.5°, 7.9±0.5°, 8.3±0.5°,8.7±0.5°, 9.0±0.5°, 10.4±0.5°, 11.2±0.5°, 11.6±0.5°, 12.3±0.5°,13.1±0.5°, 13.9±0.5°, 14.7±0.5°, 15.8±0.5°, 16.5±0.5°, 17.1±0.5°,17.9±0.5°, 18.4±0.5°, 18.9±0.5°, 19.6±0.5°, 20.4±0.5°, 21.0±0.5°,21.8±0.5°, 22.9±0.5°, 23.3±0.5°, 23.6±0.5°, 24.0±0.5°, 24.9±0.5° and26.4±0.5° (2θ, 1d.p); and/or(c) an XRPD pattern substantially as shown in FIG. 16 labelled as 1;and/or(d) having peaks at the same diffraction angles (2θ) of the XRPD patternshown in FIG. 16 labelled as 1 and optionally wherein the peaks have thesame relative intensity as the peaks shown in FIG. 16 labelled as 1;and/or(e) having major peaks at diffraction angles (2θ) and intensities asthose shown in the XRPD pattern in FIG. 16 labelled as 1; and/or(f) having major peaks as measured by XRPD at 8.7±0.5°, 17.1±0.5°,17.9±0.5° and 18.9±0.5° (2θ, 1d.p); and/or(g) interplanar spacing (d) values of 11.94±0.5 Å, 11.19±0.5 Å,10.65±0.5 Å, 10.16±0.5 Å, 9.82±0.5 Å, 8.50±0.5 Å, 7.90±0.5 Å, 7.62±0.5Å, 7.19±0.5 Å, 6.75±0.5 Å, 6.37±0.5 Å, 6.02±0.5 Å, 5.61±0.5 Å, 5.37±0.5Å, 5.18±0.5 Å, 4.95±0.5 Å, 4.82±0.5 Å, 4.69±0.5 A, 4.53±0.5 Å, 4.35±0.5Å, 4.23±0.5 Å, 4.07±0.5 Å, 3.88±0.5 Å, 3.82±0.5 Å, 3.77±0.5 Å, 3.71±0.5Å, 3.57±0.5 Å and 3.37±0.5 Å (d, 2d.p.); and/or(h) a DSC peak temperature of 174.37° C.±0.5° C. (such as 174.37°C.±0.2° C., in particular 174.37° C.±0.1° C., more particularly 174.37°C.); and/or(i) a DSC onset temperature of 171.6° C.±0.5° C. (endotherm, sharp)(such as 171.6° C.±0.2° C., in particular 171.6° C.±0.1° C., moreparticularly 171.6° C.); and/or(j) a DSC thermogram as depicted in FIG. 17 labelled as 1.

In particular, the Form C polymorph of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate provides advantages with respect to stability andcrystallinity.

Complexes

Formula (I) also includes within its scope complexes (e.g. inclusioncomplexes or clathrates with compounds such as cyclodextrins, orcomplexes with metals) of the compounds. Inclusion complexes, clathratesand metal complexes can be formed by means of methods well known to theskilled person.

Prodrugs

Also encompassed by formula (I) are any pro-drugs of the compounds ofthe formula (I). By “prodrugs” is meant for example any compound that isconverted in vivo into a biologically active compound of the formula(I).

For example, some prodrugs are esters of the active compound (e.g., aphysiologically acceptable metabolically labile ester). Duringmetabolism, the ester group (—C(═O)OR) is cleaved to yield the activedrug. Such esters may be formed by esterification, for example, of anyof the carboxylic acid groups (—C(═O)OH) in the parent compound, with,where appropriate, prior protection of any other reactive groups presentin the parent compound, followed by deprotection if required.

Examples of such metabolically labile esters include those of theformula —C(═O)OR wherein R is:

-   C₁₋₇alkyl (e.g., -Me, -Et, -nPr, -iPr, -nBu, -sBu, -iBu, -tBu);    C₁₋₇aminoalkyl (e.g., aminoethyl; 2-(N,N-diethylamino)ethyl;    2-(4-morpholino)ethyl); and acyloxy-C₁₋₇ alkyl (e.g., acyloxymethyl;    acyloxyethyl; pivaloyloxymethyl; acetoxymethyl; 1-acetoxyethyl;    1-(1-methoxy-1-methyl)ethyl-carbonxyloxyethyl; 1-(benzoyloxy)ethyl;    isopropoxy-carbonyloxymethyl; 1-isopropoxy-carbonyloxyethyl;    cyclohexyl-carbonyloxymethyl; 1-cyclohexyl-carbonyloxyethyl;    cyclohexyloxy-carbonyloxymethyl; 1-cyclohexyloxy-carbonyloxyethyl;    (4-tetrahydropyranyloxy) carbonyloxymethyl;    1-(4-tetrahydropyranyloxy)carbonyloxyethyl;    (4-tetrahydropyranyl)carbonyloxymethyl; and    1-(4-tetrahydropyranyl)carbonyloxyethyl).

Also, some prodrugs are activated enzymatically to yield the activecompound, or a compound which, upon further chemical reaction, yieldsthe active compound (for example, as in antigen-directed enzyme pro-drugtherapy (ADEPT), gene-directed enzyme pro-drug therapy (GDEPT), andligand-directed enzyme pro-drug therapy (LIDEPT), etc.). For example,the prodrug may be a sugar derivative or other glycoside conjugate, ormay be an amino acid ester derivative. In one embodiment formula (I)does not include pro-drugs of the compounds of the formula (I) withinits scope.

Advantages of Compounds of the Invention

The compounds of the formula (I) may have a number of advantages overprior art compounds.

Compounds of the invention may have particular advantage in one or moreof the following aspects:

-   -   (i) Superior selectivity versus the IKr (hERG) cardiac ion        channel;    -   (ii) Superior metabolic stability;    -   (iii) Lower P450 inhibitory liability;    -   (iv) Superior oral bioavailabilty; and/or    -   (v) Superior in vivo efficacy.

Superior Selectivity Versus the IKr (hERG) Cardiac Ion Channel

In the late 1990s a number of drugs, approved by the US FDA, had to bewithdrawn from sale in the US when it was discovered they wereimplicated in deaths caused by heart malfunction. It was subsequentlyfound that a side effect of these drugs was the development ofarrhythmias caused by the blocking of hERG channels in heart cells. ThehERG channel is one of a family of potassium ion channels the firstmember of which was identified in the late 1980s in a mutant Drosophilamelanogaster fruitfly (see Jan, L. Y. and Jan, Y. N. (1990). ASuperfamily of Ion Channels. Nature, 345(6277):672). The biophysicalproperties of the hERG potassium ion channel are described inSanguinetti, M. C., Jiang, C., Curran, M. E., and Keating, M. T. (1995).A Mechanistic Link Between an Inherited and an Acquired CardiacArrhythmia: HERG encodes the Ikr potassium channel. Cell, 81:299-307,and Trudeau, M. C., Warmke, J. W., Ganetzky, B., and Robertson, G. A.(1995). HERG, a Human Inward Rectifier in the Voltage-Gated PotassiumChannel Family. Science, 269:92-95. Therefore, elimination of hERGblocking activity remains an important consideration in the developmentof any new drug.

It has been found that many compounds of the formula (I) have reducedhERG activity and/or a good separation between IAP activity and hERGactivity (greater ‘therapeutic window’). One method for measurement ofhERG activity is the patch clamp electrophysiology method. Alternativemethods for measurement of functional hERG activity include hERG bindingassays, which can use commercially available membranes isolated fromcells stably expressing the hERG channel or commercially available celllines expressing the hERG channel.

Many compounds of the formula (I) have improved Cardiac Safety Index(CSI) [CSI=hERG IC50/Cmax(unbound)] (Shultz et al, J. Med. Chem., 2011;Redfern et al, Cardiovasc. Res., 2003). This can be due to an increasein hERG IC50 or a reduction in Cmax required for efficacy (due to betterIAP potency and/or PK).

Particular compounds of formula (I) have reduced hERG ion channelblocking activity. Particular compounds of the formula (I) have meanIC₅₀ values against hERG that are greater than 30 times, or greater than40 times, or greater than 50 times the IC₅₀ values of the compounds incellular proliferation assays. Particular compounds of the formula (I)have mean IC₅₀ values against hERG that are greater than 10 μM, moreparticularly greater than 20 μM, and more preferably greater than 30 μM.Some compounds of the invention have mean IC₅₀ values against hERG thatare greater than 40 μM or display % inhibition representative of such anIC₅₀ at concentrations of 10, 30 or 300 μM. Some compounds of theinvention have mean CSI of higher than minimum recommended value (30fold).

As can be seen from the data in Table 1 herein, the compounds ofExamples 1-34 all have a lower hERG liability than the compound Example259 (also 262 and 263) of WO 2012/143726. In particular the compounds ofExamples 1-2, 11 and 34 of the invention demonstrated an IC₅₀ of 40 μMagainst hERG whereas the compound of Example 259 (also 262 and 263) ofWO 2012/143726 displays 42% inhibition of hERG at 10 μM. Therefore,superior selectivity versus hERG is a key advantage of compounds of theinvention over prior disclosed IAP antagonist compounds, in particularthose disclosed in WO 2012/143726.

Superior Metabolic Stability

The compounds of the formula (I) may have advantageous ADMET propertiesfor example better metabolic stability (for example as determined withmouse liver microsomes), a better P450 profile and/or beneficialclearance (e.g. low clearance). These features could confer theadvantage of having more drug available in the systemic circulation toreach the appropriate site of action to exert its therapeutic effect.Increased drug concentrations to exert pharmacological action in tumourspotentially leads to improved efficacy which thereby allows reduceddosages to be administered. Thus, the compounds of formula (I) shouldexhibit reduced dosage requirements and should be more readilyformulated and administered. In addition the compound may have reducedP450 (e.g. 3A4) turnover.

Lower P450 Inhibitory Liability

Many of the compounds of the formula (I) are advantageous in that theyhave different susceptibilities to P450 enzymes. For example, theparticular compounds of the formula (I) have IC₅₀ values of greater than10 μM against each of the cytochrome P450 enzymes 1A2, 2C9, 2C19, 3A4and 2D6 (in particular 3A4). In addition particularly the compounds arenot P450 inhibitors.

Superior Oral Bioavailability

Potentially the compounds of the invention have physiochemicalproperties suitable for oral exposure (oral exposure or AUC). Inparticular, compounds of the formula (I) may exhibit improved oralbioavailability. Oral bioavailability can be defined as the ratio (F) ofthe plasma exposure of a compound when dosed by the oral route to theplasma exposure of the compound when dosed by the intravenous (i.v.)route, expressed as a percentage.

Compounds having an oral bioavailability (F value, F %) of greater than30%, more particularly greater than 40%, are particularly advantageousin that they may be adminstered orally rather than, or as well as, byparenteral administration.

Superior In Vivo Efficacy

As a result of increased potency against XIAP and/or cIAP compounds ofthe invention may have increased in vivo efficacy in cancer cell linesand in vivo models.

Methods for the Preparation of Compounds of Formula (I)

In this section, as in all other sections of this application unless thecontext indicates otherwise, references to formula (I) also include allother sub-groups and examples thereof as defined herein.

Compounds of the formula (I) can be prepared in accordance withsynthetic methods well known to the skilled person.

According to a further aspect of the invention there is provided aprocess for preparing a compound of formula (I) as hereinbefore definedwhich comprises:

(a) (i) reacting a compound of formula (II):

wherein R⁵, R⁶, U and X are as defined hereinbefore for compounds offormula (I), L¹ represents a suitable leaving group, such as a halogenatom (e.g. chlorine) and P¹ represents hydrogen or a suitable protectinggroup such as a tert-butyloxycarbonyl (tBoc) group, with a compound offormula (III):

or an optionally protected derivative thereof; wherein R¹ and R² are asdefined hereinbefore for compounds of formula (I), followed by adeprotection reaction suitable to remove the P¹ protecting group and anyother protecting groups as necessary; or

-   -   (ii) reacting a compound of formula (IV):

wherein R⁵, R⁶, X and U are as defined hereinbefore for compounds offormula (I), and L² represents a suitable leaving group such as halogen(e.g. chlorine), with a compound of formula (V):

or an optionally protected derivative thereof; wherein R¹ and R² are asdefined hereinbefore for compounds of formula (I) and P² representshydrogen or a suitable protecting group such as a tert-butyloxycarbonyl(tBoc) group, followed by a deprotection reaction suitable to remove theP² protecting group and any other protecting groups as necessary; and/or(b) deprotection of a protected derivative of a compound of formula (I);and/or(c) interconversion of a compound of formula (I) or protected derivativethereof to a further compound of formula (I) or protected derivativethereof; and(d) optional formation of a pharmaceutically acceptable salt of acompound of formula (I).

Process (a)(i) typically comprises reacting a compound of formula (II)with a compound of formula (III), optionally in the presence of asuitable additive such as potassium iodide and a suitable base such aspotassium carbonate in a suitable solvent such as acetonitrile. Such aprocess may be carried out at ambient temperature or at elevatedtemperature, e.g. 70° C.

Process (a)(ii) typically comprises reacting a compound of formula (IV)with a compound of formula (V), optionally in the presence of a suitableadditive such as potassium iodide and a suitable base such as potassiumcarbonate in a suitable solvent such as acetonitrile.

Process (b) typically comprises any suitable deprotection reaction, theconditions of which will depend upon the nature of the protecting group.When the protecting group represents tBoc, such a deprotection reactionwill typically comprise the use of a suitable acid in a suitablesolvent. For example, the acid may suitably comprise trifluoroaceticacid or hydrogen chloride and the solvent may suitably comprisedichloromethane ethyl acetate, 1,4-dioxane, methanol or water.Optionally a mixture of solvents may be used, for example aqueousmethanol or ethyl acetate/1,4-dioxane.

It will be appreciated that, when the protecting group represents tBoc,deprotection using a suitable acid as described above may generate acompound of formula (I) as a pharmaceutically acceptable salt, which maybe isolated directly. Alternatively, the compound of formula (I) may beisolated as the free base using methods well known in the art andthereafter optionally converted to a pharmaceutically acceptable saltaccording to process (d).

Process (c) typically comprises interconversion procedures known by oneskilled in the art. For example, in compounds of formula (I), a firstsubstituent may be converted by methods known by one skilled in the artinto a second, alternative substituent. A wide range of well knownfunctional group interconversions are known by a person skilled in theart for converting a precursor comound to a compound of formula I andare described in Advanced Organic Chemistry by Jerry March, 4th Edition,John Wley & Sons, 1992. For example possible metal catalysedfunctionalisations such as using organo-tin reagents (the Stillereaction), Grignard reagents and reactions with nitrogen nucleophilesare described in ‘Palladium Reagents and Catalysts’ [Jiro Tsuji, Wiley,ISBN 0-470-85032-9] and Handbook of OrganoPalladium Chemistry forOrganic Synthesis [Volume 1, Edited by Ei-ichi Negishi, Wiley, ISBN0-471-31506-0].

Process (d) may be carried out by treatment of a compound of formula (I)in the free base form, dissolved in a suitable solvent, with astoichiometric amount or an excess of a pharmaceutically acceptableorganic or inorganic acid, then isolation of the resulting salt bymethods well known in the art, e.g. evaporation of solvent orcrystallisation.

If appropriate, the reactions previously described in processes (a), (b)and (c) are followed or preceded by one or more reactions known to theskilled of the art and are performed in an appropriate order to achievethe requisite substitutions on R¹, R², R⁵ and R⁶ defined above to affordother compounds of formula (I). Non-limiting examples of such reactionswhose conditions can be found in the literature include:

-   -   protection of reactive functions,    -   deprotection of reactive functions,    -   halogenation,    -   dehalogenation,    -   dealkylation,    -   alkylation and arylation of amine, aniline, alcohol and phenol,    -   Mitsunobu reaction on hydroxyl groups,    -   cycloaddition reactions on appropriate groups,    -   reduction of nitro, esters, cyano, aldehydes,    -   transition metal-catalyzed coupling reactions,    -   acylation,    -   sulfonylation/introduction of sulfonyl groups,    -   saponification/hydrolysis of ester groups,    -   amidification or transesterification of ester groups,    -   esterification or amidification of carboxylic groups,    -   halogen exchange,    -   nucleophilic substitution with amine, thiol or alcohol,    -   reductive amination,    -   oxime formation on carbonyl and hydroxylamine groups,    -   S-oxidation,    -   N-oxidation,    -   salification.

Compounds of formula (II) may be prepared from compounds of formula (IV)in accordance with the following Scheme 1:

wherein X, U, R⁵, R⁶, L¹, L² and P¹ are as defined hereinbefore.

Step (i) of Scheme 1 typically comprises reacting the compounds offormulae (IV) and (VI), optionally in the presence of a suitableadditive such as potassium iodide and a suitable base such as potassiumcarbonate in a suitable solvent such as acetonitrile.

When L¹ represents chlorine, step (ii) of Scheme 1 typically comprisesreacting the compound of formula (VII) with a reagent capable ofconverting a hydroxyl group into a good leaving group, e.g.methylsulfonyl chloride, in the presence of a base such astriethylamine.

Compounds of formula (IV) where X represents N, U represents carbon andR⁶ represents hydroxymethyl may be prepared in accordance with thefollowing Scheme 2:

wherein L³, L⁴, L⁵ and L⁶ represent suitable leaving groups, such as ahalogen atom (i.e. fluorine, bromine or chlorine) and R⁵ and L² are asdefined hereinbefore.

When L³ and L⁴ both represent fluorine, step (i) of Scheme 2 typicallycomprises reacting a compound of formula (VIII) with a base such assodium bis(trimethylsilyl)amide in the presence of tetrahydrofuran andisobutronitrile in a suitable solvent such as toluene. An example ofsuch a reaction is shown herein in Preparation 11.

Step (ii) of Scheme 2 involves reaction with a suitable reducing agentand typically comprises reacting the compound of formula (IX) with aborane-tetrahydrofuran complex in the presence of a suitable solventsuch as tetrahydrofuran. An example of such a reaction is shown hereinin Preparation 12. Step (ii) of Scheme 2 may also typically comprisereacting the compound of formula (IX) with nickel(ii) chloridehexahydrate followed by addition of sodium borohydride. An example ofsuch a reaction is shown herein in Preparation 12, alternativeprocedure.

Step (iii) of Scheme 2 typically comprises cyclisation of the compoundof formula (X) using a suitable base e.g. potassium carbonate and anappropriate solvent such as NMP. An example of such a reaction is shownherein in Preparation 13.

Step (iv) of Scheme 2 typically comprises reacting the compound offormula (XI) with a compound of formula R⁵-M, wherein R⁵ is as definedhereinbefore and M represents the residue of an organometallic speciessuch that R⁵-M represents a nucleophilic organometallic reagent such asan organozinc halide. Step (iv) typically also comprises the use oflithium bromide, a catalyst, such as[1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II)dichloride, in a suitable solvent system e.g. tetrahydrofuran and NMP.An example of such a reaction is shown herein in Preparation 15.

Step (v) of Scheme 2 typically comprises halogenation of the compound offormula (XII) for example using N-bromosuccinimide in dimethylformamide.An example of such a reaction is shown herein in Preparation 16.

Step (vi) of Scheme 2 involves lithiation and reaction with a suitableelectrophile for introduction of the formyl group, and typicallycomprises reacting the compound of formula (XIII) with MeLi in THFfollowed by addition of tBuLi in hexane followed by addition ofdimethylformamide. An example of such a reaction is shown herein inPreparation 17.

Step (vii) of Scheme 2 involves reduction of the formyl group with asuitable reducing agent and typically comprises reacting the compound offormula (XIV) with sodium borohydride in methanol. An example of such areaction is shown herein in Preparation 17.

When L² represents a halogen such as chlorine, step (viii) of Scheme 2typically comprises reacting the compound of formula (XV) with ahaloacetyl halide such as chloroacetyl chloride in MeCN followed byaddition of potassium carbonate in methanol. An example of such areaction is shown herein in Preparation 18. Alternatively, compounds offormula (XIII) may be converted to compounds of formula (XV) byfollowing a sequence analogous to that described in Preparations 25-29inclusive.

It will be appreciated that compounds of formula (XV) where R⁶represents CH(OR^(x))CH₂OR^(z) may be prepared in an analogous manner toScheme 2 above by varying steps (v) onwards of Scheme 2. Examples ofsuitable reaction sequences are shown herein in Preparations 38-42.

Compounds where X represents NR³, U represents carbon and R⁶ is ═O canbe synthesised using functional group interconversions on appropriateintermediates of Scheme 2 or protected derivatives thereof, for exampleas demonstrated in Preparations 22-24, 30-35 and 50.

It will also be appreciated that compounds of formula (IV) where R⁵represents unsubstituted n-butyl or alternatively substituted benzylgroups may be prepared in an analogous manner to Scheme 2 above byvarying the organometallic reagent used in step (iv) of Scheme 2. Anexample of such a reaction is shown herein in Preparations 15A, 15B and15C.

Compounds where X represents CR⁴, U represents nitrogen and R⁶represents oxo can be synthesised using analogous sequences to thosedescribed in preparations 43-49 and 51-58.

Compounds of formula (V), or optionally protected derivatives thereof,may be prepared in accordance with the following Scheme 3:

wherein R¹, R² and P² are as defined hereinbefore for compounds offormula (V), L⁷ represents a suitable leaving group such as a halogenatom (e.g. chlorine) and P³ represents a suitable protecting group, suchas benzyl.

When P³ represents benzyl, step (i) of Scheme 3 typically comprisesreacting a compound of formula (XVI) with benzaldehyde in the presenceof a suitable reducing agent such as sodium triacetoxyborohydride and1,2-dichloroethane. An example of such a reaction is shown herein inPreparation 5.

When L⁷ represents chlorine, step (ii) of Scheme 3 typically comprisesreacting a compound of formula (XVII) with methanesulfonyl chloride inthe presence of triethylamine and dichloromethane. An example of such areaction is shown herein in Preparation 6.

Step (iii) of Scheme 3 typically comprises reacting the compounds offormulae (XVIII) and (XIX) in the presence of a base such as potassiumcarbonate, an additive such as potassium iodide in a suitable solventsuch as acetonitrile. An example of such a reaction is shown herein inPreparation 7.

Step (iv) of Scheme 3 typically comprises a deprotection reaction. Forexample, when P³ represents benzyl, step (iv) typically compriseshydrogenation of the compound of formula (XX) in the presence of asuitable catalyst such as palladium on carbon in a suitable solventsystem such as ethanol or a mixture of acetic acid and ethanol. Anexample of such a reaction is shown herein in Preparation 8.

Alternatively compounds of formula (I) can be synthesised by reacting acompound of formula (XXI):

or an optionally protected derivative thereof, wherein R¹ and R² are asdefined hereinbefore for compounds of formula (I) and P² represents asuitable protecting group such as a tert-butyloxycarbonyl (tBoc) group,with a compound of formula (XXII):

wherein X, U, R⁵ and R⁶ are as defined hereinbefore followed by adeprotection reaction suitable to remove the protecting group P² and anyadditional protecting groups.

One example of a suitable compound of formula (XXII) includes a compoundof formula (XV) as defined hereinbefore.

This reaction typically comprises reacting a compound of formula (XXI)with a compound of formula (XXII), such as a compound of formula (XV),in a suitable solvent and at a suitable temperature e.g. ambienttemperature, in the presence of a suitable base and a reagent capable ofactivating the carboxylic acid group present in the compound of formula(XXI). A suitable solvent should be inert toward the reagents used, forexample dichloromethane. Examples of suitable bases are triethylamineand N,N-diisopropylethylamine (DIPEA). Examples of suitable activatingreagents are bromo-tris-pyrrolidino-phosphonium hexofluorophosphate(PyBrop), O-benzotriazole-N, N, N′,N′-tetramethyl-uronium-hexafluoro-phosphate (HBTU),1,1′-carbonyldiimidazole,1-ethyl-3-(3′-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate) (HATU). This process may optionally be carried outin the presence of a catalytic or stoichiometric amount of a suitableco-activating reagent such as 1-hydroxybenzotriazole (HOBt) or1-hydroxyazabenzotriazole (HOAt).

Compounds of formula (XXI) or optionally protected derivatives thereofmay be prepared from compounds of formula (V) or optionally protectedderivatives thereof as defined above by methods well known in the art,for example by reaction with an ester of a monohaloacetic acid such asbenzyl bromoacetate in the presence of a suitable base such as potassiumcarbonate in a suitable solvent such as acetonitrile; and subsequentester hydrolysis (or optionally hydrogenolysis in the case of a benzylester). Compounds of formula (I) may be prepared following an analogoussequence to that described in Preparations 1-5.

Compounds of formula (XXII) may be prepared by using analogous sequencesto those described in Scheme 2 or the following preparations: 38-42;22-24, 30-35 and 50; or 43-49 and 51-58.

It will be appreciated that certain compounds e.g. compounds of formulae(I), (II), (III), (V), (VI), (VII), (XVI), (XVII), (XVIII), (XIX), (XX),(XXI) and (XXII) can exist in different diastereomeric and/orenantiomeric forms and that processes for their preparation may make useof enantiomerically pure synthetic precursors.

Alternatively racemic precursors may be used and the mixtures ofdiastereoisomers generated in these process may be separated by methodswell known to the person skilled in the art, for example usingnon-chiral or chiral preparative chromatography or resolution usingdiastereomeric derivatives: for example crystallisation of a salt formedwith an enantiomerically pure acid such as L-tartaric acid; orenantiomer separation of a diastereomeric derivative formed bycovalently linking a enantiomerically pure chiral auxiliary onto thecompound, followed by separation using conventional methods such aschiral chromatography. The aforementioned covalent linkage is thencleaved to generate the appropriate enantiomerically pure product.

The required intermediates, for example compounds of formula (III),(VI), (VIII), R⁵-M, (XVI) and (XIX) are either commercially available,known in the literature, prepared by methods analogous to those in theliterature or prepared by methods analogous to those described in theexample experimental procedures below. Other compounds may be preparedby functional group interconversion of the groups R¹, R², R⁵ and R⁶using methods well known in the art.

In a further embodiment the invention provides a novel intermediate. Inone embodiment the invention provides a novel intermediate of formula(II) or (IV) or (V) or (VII) or (XX). In an alternative embodiment theinvention provides a novel intermediate of formula (XXI) or (XXII).

Protecting Groups

In many of the reactions described above, it may be necessary to protectone or more groups to prevent reaction from taking place at anundesirable location on the molecule. Examples of protecting groups, andmethods of protecting and deprotecting functional groups, can be foundin Protective Groups in Organic Synthesis (T. Green and P. Wuts; 3rdEdition; John Wiley and Sons, 1999).

In particular the groups R¹ and R² may be synthesised in protected formsand the protecting groups removed to generate a compound of formula (I).

A hydroxy group may be protected, for example, as an ether (—OR) or anester (—OC(═O)R), for example, as: a t-butyl ether; a tetrahydropyranyl(THP) ether; a benzyl, benzhydryl (diphenylmethyl), or trityl(triphenylmethyl) ether; a trimethylsilyl or t-butyldimethylsilyl ether;or an acetyl ester (—OC(═O)CH₃).

An aldehyde or ketone group may be protected, for example, as an acetal(R—CH(OR)₂) or ketal (R₂C(OR)₂), respectively, in which the carbonylgroup (>C═O) is treated with, for example, a primary alcohol. Thealdehyde or ketone group is readily regenerated by hydrolysis using alarge excess of water in the presence of acid.

An amine group may be protected, for example, as an amide (—NRCO—R) or acarbamate (—NRCO—OR), for example, as: a methyl amide (—NHCO—CH₃); abenzyl carbamate (—NHCO—OCH₂C₆H₅, —NH-Cbz or NH—Z); as a t-butylcarbamate (—NHCO—OC(CH₃)₃, —NH-Boc); a 2-biphenyl-2-propyl carbamate(—NHCO—OC(CH₃)₂C₆H₄C₆H₅, —NH-Bpoc), as a 9-fluorenylmethyl carbamate(—NH—Fmoc), as a 6-nitroveratryl carbamate (—NH—Nvoc), as a2-trimethylsilylethyl carbamate (—NH-Teoc), as a 2,2,2-trichloroethylcarbamate (—NH-Troc), as an allyl carbamate (—NH-Alloc), or as a2(-phenylsulfonyl)ethyl carbamate (—NH—Psec).

For example, in compounds of formula II contains an amino group, theamino group can be protected by means of a protecting group ashereinbefore defined, one particular group being thetert-butyloxycarbonyl (Boc) group while the additionalfunactionalisation is introduced. Where no subsequent modification ofthe amino group is required, the protecting group can be carried throughthe reaction sequence to give an N-protected form of a compound of theformula (I) which can then be de-protected by standard methods (e.g.treatment with acid in the case of the Boc group) to give the compoundof formula (I).

Other protecting groups for amines, such as cyclic amines andheterocyclic N—H groups, include toluenesulfonyl (tosyl) andmethanesulfonyl (mesyl) groups, benzyl groups such as apara-methoxybenzyl (PMB) group and tetrahydropyranyl (THP) groups.

A carboxylic acid group may be protected as an ester for example, as: anC₁₋₇ alkyl ester (e.g., a methyl ester; a t-butyl ester); a C₁₋₇haloalkyl ester (e.g., a C₁₋₇ trihaloalkyl ester); a triC₁₋₇alkylsilyl-C₁₋₇alkyl ester; or a C₅₋₂₀ aryl-C₁₋₇ alkyl ester (e.g., abenzyl ester; a nitrobenzyl ester; para-methoxybenzyl ester. A thiolgroup may be protected, for example, as a thioether (—SR), for example,as: a benzyl thioether; an acetamidomethyl ether (—S—CH₂NHC(═O)CH₃).

Isolation and Purification of the Compounds of the Invention

The compounds of the invention can be isolated and purified according tostandard techniques well known to the person skilled in the art andexamples of such methods include chromatographic techniques such ascolumn chromatography (e.g. flash chromatography) and HPLC. Onetechnique of particular usefulness in purifying the compounds ispreparative liquid chromatography using mass spectrometry as a means ofdetecting the purified compounds emerging from the chromatographycolumn.

Preparative LC-MS is a standard and effective method used for thepurification of small organic molecules such as the compounds describedherein. The methods for the liquid chromatography (LC) and massspectrometry (MS) can be varied to provide better separation of thecrude materials and improved detection of the samples by MS.Optimisation of the preparative gradient LC method will involve varyingcolumns, volatile eluents and modifiers, and gradients. Methods are wellknown in the art for optimising preparative LC-MS methods and then usingthem to purify compounds. Such methods are described in Rosentreter U,Huber U.; Optimal fraction collecting in preparative LC/MS; J CombChem.; 2004; 6(2), 159-64 and Leister W, Strauss K, Wisnoski D, Zhao Z,Lindsley C., Development of a custom high-throughput preparative liquidchromatography/mass spectrometer platform for the preparativepurification and analytical analysis of compound libraries; J CombChem.; 2003; 5(3); 322-9. An example of such a system for purifyingcompounds via preparative LC-MS is described below in the Examplessection of this application (under the heading “Mass DirectedPurification LC-MS System”).

Methods of recrystallisation of compounds of formula (I) and saltthereof can be carried out by methods well known to the skilledperson—see for example (P. Heinrich Stahl (Editor), Camille G. Wermuth(Editor), ISBN: 3-90639-026-8, Handbook of Pharmaceutical Salts:Properties, Selection, and Use, Chapter 8, Publisher Wiley-VCH).Products obtained from an organic reaction are seldom pure when isolateddirectly from the reaction mixture. If the compound (or a salt thereof)is solid, it may be purified and/or crystallized by recrystallisationfrom a suitable solvent. A good recrystallisation solvent shoulddissolve a moderate quantity of the substance to be purified at elevatedtemperatures but only a small quantity of the substance at lowertemperature. It should dissolve impurities readily at low temperaturesor not at all. Finally, the solvent should be readily removed from thepurified product. This usually means that it has a relatively lowboiling point and a person skilled in the art will know recrystallisingsolvents for a particular substance, or if that information is notavailable, test several solvents. To get a good yield of purifiedmaterial, the minimum amount of hot solvent to dissolve all the impurematerial is used. In practice, 3-5% more solvent than necessary is usedso the solution is not saturated. If the impure compound contains animpurity which is insoluble in the solvent it may then be removed byfiltration and then allowing the solution to crystallize. In addition,if the impure compound contains traces of coloured material that are notnative to the compound, it may be removed by adding a small amount ofdecolorizing agent e.g. activating charcoal to the hot solution,filtering it and then allowing it to crystallize. Usuallycrystallization spontaneously occurs upon cooling the solution. If it isnot, crystallization may be induced by cooling the solution below roomtemperature or by adding a single crystal of pure material (a seedcrystal). Recrystallisation can also be carried out and/or the yieldoptimized by the use of an anti-solvent or co-solvent. In this case, thecompound is dissolved in a suitable solvent at elevated temperature,filtered and then an additional solvent in which the required compoundhas low solubility is added to aid crystallization. The crystals arethen typically isolated using vacuum filtration, washed and then dried,for example, in an oven or via desiccation.

Other examples of methods for purification include sublimation, whichincludes an heating step under vacuum for example using a cold finger,and crystallization from melt (Crystallization Technology Handbook 2ndEdition, edited by A. Mersmann, 2001).

Biological Effects

The compounds of the invention, subgroups and examples thereof, areantagonists of inhibitor of apoptosis protein (IAP), and which may beuseful in preventing or treating disease states or conditions describedherein. In addition the compounds of the invention, and subgroupsthereof, will be useful in preventing or treating diseases or conditionmediated by IAP. References to the preventing or prophylaxis ortreatment of a disease state or condition such as cancer include withintheir scope alleviating or reducing the incidence of cancer.

Thus, for example, it is envisaged that the compounds of the inventionwill be useful in alleviating or reducing the incidence of cancer.

The compounds of the present invention may be useful for the treatmentof the adult population. The compounds of the present invention may beuseful for the treatment of the pediatric population.

More particularly, the compounds of the formula (I) and sub-groupsthereof are antagonists of IAP. For example, compounds of the inventionhave affinity against XIAP, cIAP1 and/or cIAP2, and in particular an IAPselected from XIAP and cIAP1.

Particular compounds are compounds that have affinity for one or moreIAP selected from XIAP, cIAP1 and cIAP2. Particular compounds of theinvention are those having IC₅₀ values of less than 0.1 μM.

The antagonist compounds of formula (I) are capable of binding to IAPand exhibiting potency for IAP. In one embodiment the antagonistcompounds of formula (I) exhibit selectivity for one or more IAP overother IAP family members, and may be capable of binding to and/orexhibiting affinity for XIAP and/or cIAP in preference to binding toand/or exhibiting affinity for other of the IAP family members.

In addition many of the compounds of the invention exhibit selectivityfor the XIAP compared to cIAP or vice versa, selectivity for the cIAPcompared to XIAP (in particular cIAP1), and such compounds represent oneembodiment of the invention. In particular compounds of the inventionmay have at least 10 times greater affinity against one or more IAPfamily member in particular XIAP, cIAP1 and/or cIAP2 than other IAPfamily members. This can be determined using the methods describedherein. In a further embodiment compounds of the invention may haveequivalent affinity for XIAP, cIAP1 and/or cIAP2, in particularequivalent affinity (i.e. less than 10-fold difference in affinity) forXIAP and cIAP1.

Activity against XIAP and cIAP1 may be particularly advantageous.Antagonising XIAP and cIAP1 with equipotency should enable triggering ofapoptosis via activation of caspase-8 and the switch away frompro-survival NF-κB signalling towards apoptosis; and potent antagonismof XIAP will ensure that apoptosis is achieved before any inherentresistance mechanism is upregulated to block the process. On depletionof cIAP1 via autoubiquitination and proteasomal degradation there is atemporary upregulation of NF-κB signalling that is responsible forexpression of TNF-alpha in sensitive cell lines—this is also responsiblefor upregulation of anti-apoptotic factors such as cIAP2 and c-FLIP.Hence the need for potent XIAP antagonism to potentiate effector caspaseactivation and cell death, rather than allowing cIAP2-mediatedresistance to build up. It is generally believed that toxicities thatarise on dosing these compounds in vivo will arise from the temporaryinduction of NF-κB signalling and resultant upregulation ofpro-inflammatory cytokines, which is mediated solely by cIAP1/2antagonism. Therefore dual potency should enable a therapeutic window tobe achieved before dose-limiting toxicities are encountered.

IAP function in controlling programmed cell death has also beenimplicated in many diseases, including disorders associated with cellaccumulation (e.g. cancer, autoimmune disorders, inflammation andrestenosis), disorders where excessive apoptosis results in cell loss(e.g. stroke, heart failure, neurodegeneration such as Alzheimers'disease, Parkinson's disease, Huntington's disease, amyotrophic lateralsclerosis, AIDS, ischemia (stroke, myocardial infarction) andosteoporosis or treating autoimmune diseases such as multiple sclerosis(MS).

Therefore, it is also envisaged that the compounds of the invention maybe useful in treating other conditions such as inflammation (for examplearthritis including rheumatoid arthririts), hepatitis, ulcerativecolitis, gastritis, autoimmunity, restenosis, stroke, heart failure,neurodegenerative conditions such as Alzheimers' disease, Parkinson'sdisease, Huntington's disease, myotonic dystrophy, and amyotrophiclateral sclerosis, AIDS, ischemia such as traumatic brain injury, spinalcord injury, cerebral ischemia, cerebral ischemia/reperfusion (I/R)injury, acute and chronic CNS injury ischemia, stroke or myocardialinfarction, degenerative diseases of the musculoskeletal system such asosteoporosis, autoimmune diseases such as multiple sclerosis (MS) andType I diabetes, and eye diseases such as retinal degeneration whichresult from loss of control of programmed cell death. In one embodimentthe compounds of the invention may be useful in treating viralinfections such as herpes virus, pox virus, Epstein-Barr virus, Sindbisvirus, adenovirus, HIV, HPV, hepititis for example hepatitis B (HBV) orhepatitis C (HCV) and HCMV or in mycobacterial infections such astuberculosis (TB).

As a consequence of their affinity for IAP, the compounds will be usefulin providing a means of controlling programmed cell death. It istherefore anticipated that the compounds may prove useful in treating orpreventing proliferative disorders such as cancers. In addition, thecompounds of the invention may be useful in the treatment of diseases inwhich there is a disorder associated with cell accumulation or whereexcessive apoptosis results in cell loss.

Examples of cancers (and their benign counterparts) which may be treated(or inhibited) include, but are not limited to tumours of epithelialorigin (adenomas and carcinomas of various types includingadenocarcinomas, squamous carcinomas, transitional cell carcinomas andother carcinomas) such as carcinomas of the bladder and urinary tract,breast, gastrointestinal tract (including the esophagus, stomach(gastric), small intestine, colon, rectum and anus), liver(hepatocellular carcinoma), gall bladder and biliary system, exocrinepancreas, kidney, lung (for example adenocarcinomas, small cell lungcarcinomas, non-small cell lung carcinomas, bronchioalveolar carcinomasand mesotheliomas), head and neck (for example cancers of the tongue,buccal cavity, larynx, pharynx, nasopharynx, tonsil, salivary glands,nasal cavity and paranasal sinuses), ovary, fallopian tubes, peritoneum,vagina, vulva, penis, cervix, myometrium, endometrium, thyroid (forexample thyroid follicular carcinoma), adrenal, prostate, skin andadnexae (for example melanoma, basal cell carcinoma, squamous cellcarcinoma, keratoacanthoma, dysplastic naevus); haematologicalmalignancies (i.e. leukemias, lymphomas) and premalignant haematologicaldisorders and disorders of borderline malignancy includinghaematological malignancies and related conditions of lymphoid lineage(for example acute lymphocytic leukemia [ALL], chronic lymphocyticleukemia [CLL], B-cell lymphomas such as diffuse large B-cell lymphoma[DLBCL], follicular lymphoma, Burkitt's lymphoma, mantle cell lymphoma,T-cell lymphomas and leukaemias, natural killer [NK] cell lymphomas,Hodgkin's lymphomas, hairy cell leukaemia, monoclonal gammopathy ofuncertain significance, plasmacytoma, multiple myeloma, andpost-transplant lymphoproliferative disorders), and haematologicalmalignancies and related conditions of myeloid lineage (for exampleacute myelogenous leukemia [AML], chronic myelogenous leukemia [CML],chronic myelomonocytic leukemia [CMML], hypereosinophilic syndrome,myeloproliferative disorders such as polycythaemia vera, essentialthrombocythaemia and primary myelofibrosis, myeloproliferative syndrome,myelodysplastic syndrome, and promyelocytic leukemia); tumours ofmesenchymal origin, for example sarcomas of soft tissue, bone orcartilage such as osteosarcomas, fibrosarcomas, chondrosarcomas,rhabdomyosarcomas, leiomyosarcomas, liposarcomas, angiosarcomas,Kaposi's sarcoma, Ewing's sarcoma, synovial sarcomas, epithelioidsarcomas, gastrointestinal stromal tumours, benign and malignanthistiocytomas, and dermatofibrosarcoma protuberans; tumours of thecentral or peripheral nervous system (for example astrocytomas, gliomasand glioblastomas, meningiomas, ependymomas, pineal tumours andschwannomas); endocrine tumours (for example pituitary tumours, adrenaltumours, islet cell tumours, parathyroid tumours, carcinoid tumours andmedullary carcinoma of the thyroid); ocular and adnexal tumours (forexample retinoblastoma); germ cell and trophoblastic tumours (forexample teratomas, seminomas, dysgerminomas, hydatidiform moles andchoriocarcinomas); and paediatric and embryonal tumours (for examplemedulloblastoma, neuroblastoma, Wilms tumour, and primitiveneuroectodermal tumours); or syndromes, congenital or otherwise, whichleave the patient susceptible to malignancy (for example XerodermaPigmentosum).

Growth of cells is a closely controlled function. Cancer, a condition ofabnormal cell growth, results when cells replicate in an uncontrolledmanner (increasing in number), uncontrollably grow (getting larger)and/or experience reduced cell death by apoptosis (programmed celldeath), necrosis, or annoikis. In one embodiment abnormal cell growth isselected from uncontrolled cell proliferation, excessive cell growth orreduced programmed cell death. In particular, the condition or diseaseof abnormal cell growth is a cancer. Thus, in the pharmaceuticalcompositions, uses or methods of this invention for treating a diseaseor condition comprising abnormal cell growth (i.e. uncontrolled and/orrapid cell growth), the disease or condition comprising abnormal cellgrowth in one embodiment is a cancer.

In one embodiment the haematological malignancies is leukaemia. Inanother embodiment the haematological malignancies is lymphoma.

In one embodiment the disease to be treated is leukaemia, such as acuteand chronic leukaemias, acute myeloid leukaemia (AML), and chroniclymphocytic leukaemia (CLL). In one embodiment the leukaemia isrefractory DLBCL.

In one embodiment the lymphoma is MALT lymphoma. In one embodiment theleukemia is AML.

In one embodiment the haematological malignancy is multiple myeloma.

Many diseases are characterized by persistent and unregulatedangiogenesis. Chronic proliferative diseases are often accompanied byprofound angiogenesis, which can contribute to or maintain aninflammatory and/or proliferative state, or which leads to tissuedestruction through the invasive proliferation of blood vessels. Tumourgrowth and metastasis have been found to be angiogenesis-dependent.Compounds of the invention may therefore be useful in preventing anddisrupting initiation of tumour angiogenesis. In particular, thecompounds of the invention may be useful in the treatment of metastasisand metastatic cancers.

Metastasis or metastatic disease is the spread of a disease from oneorgan or part to another non-adjacent organ or part. The cancers whichcan be treated by the compounds of the invention include primary tumours(i.e. cancer cells at the originating site), local invasion (cancercells which penetrate and infiltrate surrounding normal tissues in thelocal area), and metastatic (or secondary) tumours ie. tumours that haveformed from malignant cells which have circulated through thebloodstream (haematogenous spread) or via lymphatics or across bodycavities (trans-coelomic) to other sites and tissues in the body.

Particular cancers include hepatocellular carcinoma, melanoma,oesophageal, renal, colon, colorectal, lung e.g. mesothelioma or lungadenocarcinoma, breast, bladder, gastrointestinal, ovarian and prostatecancers.

Particular cancers include renal, melanoma, colon, lung, breast, ovarianand prostate cancers. In one embodiment the cancer is selected frommelanoma, colon, breast and ovarian. In one embodiment the cancer ismelanoma. In one embodiment the cancer is infammatory breast cancer.

In one embodiment the cancer is lung cancer, for example mesotheliomaincluding malignant peritoneal mesothelioma or malignant pleuralmesothelioma.

In one embodiment the cancer is breast cancer, in particular triplenegative (triple −ve) breast cancer.

In one embodiment the cancer is colorectal cancer.

A further aspect of the invention includes a compound of the inventionfor use in the prophylaxis or treatment of cancer in a patient selectedfrom a sub-population possessing cancers with a high inflammatorycomponent. Such cancers are also known as “inflammatory phenotype” andinclude tumours with elevated cytokine signalling (e.g. TNF). In oneembodiment the cancer is an inflammatory tumour, for example, melanoma,colon, breast and ovarian, in particular, melanoma.

In one embodiment the melanoma is ras mutant melanoma.

Certain cancers are resistant to treatment with particular drugs. Thiscan be due to the type of the tumour (most common epithelialmalignancies are inherently chemoresistant) or resistance can arisespontaneously as the disease progresses or as a result of treatment. Inthis regard, references to mesothelioma includes mesothelioma withresistance towards topoisomerase poisons, alkylating agents,antitubulines, antifolates, platinum compounds and radiation therapy, inparticular cisplatin-resistant mesothelioma. Similarly references tomultiple myeloma includes bortezomib-sensitive multiple myeloma orrefractory multiple myeloma and references to chronic myelogenousleukemia includes imitanib-sensitive chronic myelogenous leukemia andrefractory chronic myelogenous leukemia.

The cancers may be cancers which are sensitive to antagonism of any oneor more IAP selected from XIAP, cIAP1, cIAP2, NAIP, ILP2, ML-IAP,survivin and BRUCE, more particularly XIAP, cIAP1, cIAP2, ML-IAP, mostparticularly XIAP.

It is further envisaged that the compounds of the invention, and inparticular those compounds having IAP affinity will be particularlyuseful in the treatment or prevention of cancers of a type associatedwith or characterised by the presence of elevated levels of IAP oramplification of 11q22 for example the cancers referred to in thiscontext in the introductory section of this application.

Elevated levels of IAP due to overexpression of IAP is found in manycancers and is associated with a poor prognosis. In addition, cancerswith the 11q22 amplification may also be sensitive to an IAP antagonist.The elevated levels of IAP and amplification of 11q22 can be identifiedby the techniques outlined herein. Whether a particular cancer is onewhich is sensitive to IAP function, may be determined by a method as setout in the section headed “Methods of Diagnosis”.

A further aspect provides the use of a compound for the manufacture of amedicament for the treatment of a disease or condition as decribedherein, in particular cancer.

The compounds may also be useful in the treatment of tumour growth,pathogenesis, resistance to chemo- and radio-therapy by sensitisingcells to chemotherapy and as an anti-metastatic agent.

Therapeutic anticancer interventions of all types necessarily increasethe stresses imposed on the target tumour cells. In mitigating thedeleterious effects of such stresses, IAPs are directly implicated inresisting the effects of cancer drugs and treatment regimens. Thus,antagonists of IAP represent a class of chemotherapeutics with thepotential for: (i) sensitizing malignant cells to anticancer drugsand/or treatments; (ii) alleviating or reducing the incidence ofresistance to anticancer drugs and/or treatments; (iii) reversingresistance to anticancer drugs and/or treatments; (iv) potentiating theactivity of anticancer drugs and/or treatments; (v) delaying orpreventing the onset of resistance to anticancer drugs and/ortreatments.

As a consequence of their affinity for IAP, the compounds will be usefulin providing a means of controlling programmed cell death. Therefore, itis also envisaged that the compounds of the invention may be useful intreating other conditions such as inflammatory disorders such ashepatitis, ulcerative colitis, and gastritis; neurodegenerativeconditions such as Alzheimers' disease, Parkinson's disease,Huntington's disease, myotonic dystrophy, and amyotrophic lateralsclerosis; AIDS, ischemia such as restenosis, traumatic brain injury,spinal cord injury, cerebral ischemia, cerebral ischemia/reperfusion(I/R) injury, acute and chronic CNS injury ischemia, stroke ormyocardial infarction; degenerative diseases of the musculoskeletalsystem such as osteoporosis; autoimmune diseases such as multiplesclerosis (MS) and Type I diabetes, and eye diseases such as retinaldegeneration.

The affinity of the compounds of the invention as antagonists of IAP canbe measured using the biological and biophysical assays set forth in theexamples herein and the level of affinity exhibited by a given compoundcan be defined in terms of the IC₅₀ value. Particular compounds of thepresent invention are compounds having an IC₅₀ value of less than 1 μM,more particularly less than 0.1 μM.

In one embodiment the invention provides a compound for use in thetreatment of a disease or condition which is mediated by IAP (e.g. XIAPand/or cIAP e.g. cIAP1). In a further embodiment the invention providesa compound for use in the treatment of a disease or condition whichoverexpresses IAP (e.g. XIAP and/or cIAP e.g. cIAP1).

In one embodiment the invention provides a compound for use in thetreatment of a disease or condition which is mediated by IAP, whereinthe compound is an antagonist of IAP having an IC₅₀ of less than 50 μMin at least one assay (e.g. a displacement binding) against an IAP. Inparticular the IAP is XIAP, cIAP1 and/or cIAP2. In a further embodimentthe disease or condition which is mediated by IAP is a cancer which ischaracterised by overexpression of at least one IAP and/or amplicationof 11q22.

In one embodiment the invention provides a compound for use in thetreatment of a disease or condition which is mediated by IAP, whereinthe compound has an IC₅₀ of less than 10 μM against at least one IAP inan assay (e.g. displacement binding) against IAP.

A further aspect provides the use of a compound for the manufacture of amedicament for the treatment of a disease or condition which is mediatedby IAP, wherein the compound is an antagonist of IAP having an IC₅₀ ofless than 50 μM against at least one IAP in an assay (e.g. adisplacement binding).

Methods of Diagnosis

Prior to administration of a compound of the formula (I), a patient maybe screened to determine whether a disease or condition from which thepatient is or may be suffering is one which would be susceptible totreatment with a compound having affinity for IAP. The term ‘patient’includes human and veterinary subjects.

For example, a biological sample taken from a patient may be analysed todetermine whether a condition or disease, such as cancer, that thepatient is or may be suffering from is one which is characterised by agenetic abnormality or abnormal protein expression which leads toup-regulation of the levels of IAP or to sensitisation of a pathway tonormal IAP function or to upregulation of a biochemical pathwaydownstream of IAP activation.

Examples of such abnormalities that result in activation orsensitisation of the IAP, loss of, or inhibition of apoptotic pathways,up-regulation of the receptors or ligands, cytogenetic aberrations orpresence of mutant variants of the receptors or ligands. Tumours withup-regulation of IAP, in particular over-expression of IAP, may beparticularly sensitive to IAP antagonists. For example, overexpressionof XIAP and cIAP has been identified in a range of cancers as discussionin the Background section.

Amplification of chromosome 11q22 has been detected in cell lines andprimary tumours from squamous cell carcinomas of the esophagus (Imoto etal., 2001) and cervix (Imoto et al., 2002) as well as in primary lungcancers/cell lines (Dai et al., 2003). Immunohistochemistry and westernblot analysis have identified cIAP1 and cIAP2 as potential oncogenes inthis region as both are overexpressed in cancers in which this rareamplification arises.

The term up-regulation includes elevated expression or over-expression,including gene amplification (i.e. multiple gene copies), cytogeneticaberration and increased expression by a transcriptional effect. Thus,the patient may be subjected to a diagnostic test to detect a markercharacteristic of up-regulation of IAP. The term diagnosis includesscreening. By marker we include genetic markers including, for example,the measurement of DNA composition to identify presence of mutations ofIAP or 11q22 amplification. The term marker also includes markers whichare characteristic of up regulation of IAP, including protein levels,protein state and mRNA levels of the aforementioned proteins.

The diagnostic tests and screens are typically conducted on a biologicalsample (i.e. body tissue or body fluids) selected from tumour biopsysamples, blood samples (isolation and enrichment of shed tumour cells),cerebrospinal fluid, plasma, serum, saliva, stool biopsies, sputum,chromosome analysis, pleural fluid, peritoneal fluid, buccal spears,skin biopsy or urine.

Methods of identification and analysis of cytogenetic aberration,genetic amplification, mutations and up-regulation of proteins are knownto a person skilled in the art. Screening methods could include, but arenot limited to, standard methods such as reverse-transcriptasepolymerase chain reaction (RT-PCR) or in situ hybridization such asfluorescence in situ hybridization (FISH).

In screening by RT-PCR, the level of mRNA in the tumour is assessed bycreating a cDNA copy of the mRNA followed by amplification of the cDNAby PCR. Methods of PCR amplification, the selection of primers, andconditions for amplification, are known to a person skilled in the art.Nucleic acid manipulations and PCR are carried out by standard methods,as described for example in Ausubel, F. M. et al., eds. (2004) CurrentProtocols in Molecular Biology, John Wiley & Sons Inc., or Innis, M. A.et al., eds. (1990) PCR Protocols: a guide to methods and applications,Academic Press, San Diego. Reactions and manipulations involving nucleicacid techniques are also described in Sambrook et al., (2001), 3rd Ed,Molecular Cloning: A Laboratory Manual, Cold Spring Harbor LaboratoryPress. Alternatively a commercially available kit for RT-PCR (forexample Roche Molecular Biochemicals) may be used, or methodology as setforth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659,5,272,057, 5,882,864, and 6,218,529 and incorporated herein byreference.

An example of an in situ hybridisation technique for assessing mRNAexpression would be fluorescence in situ hybridisation (FISH) (seeAngerer (1987) Meth. Enzymol., 152: 649).

Generally, in situ hybridization comprises the following major steps:(1) fixation of tissue to be analyzed; (2) prehybridization treatment ofthe sample to increase accessibility of target nucleic acid, and toreduce nonspecific binding; (3) hybridization of the mixture of nucleicacids to the nucleic acid in the biological structure or tissue; (4)post-hybridization washes to remove nucleic acid fragments not bound inthe hybridization, and (5) detection of the hybridized nucleic acidfragments. The probes used in such applications are typically labelled,for example, with radioisotopes or fluorescent reporters. Particularprobes are sufficiently long, for example, from about 50, 100, or 200nucleotides to about 1000 or more nucleotides, to enable specifichybridization with the target nucleic acid(s) under stringentconditions. Standard methods for carrying out FISH are described inAusubel, F. M. et al., eds. (2004) Current Protocols in MolecularBiology, John Wiley & Sons Inc and Fluorescence In Situ Hybridization:Technical Overview by John M. S. Bartlett in Molecular Diagnosis ofCancer, Methods and Protocols, 2nd ed.; ISBN: 1-59259-760-2; March 2004,pps. 077-088; Series: Methods in Molecular Medicine.

Methods for gene expression profiling are described by (DePrimo et al.(2003), BMC Cancer, 3:3). Briefly, the protocol is as follows:double-stranded cDNA is synthesized from total RNA using a (dT)24oligomer for priming first-strand cDNA synthesis, followed by secondstrand cDNA synthesis with random hexamer primers. The double-strandedcDNA is used as a template for in vitro transcription of cRNA usingbiotinylated ribonucleotides. cRNA is chemically fragmented according toprotocols described by Affymetrix (Santa Clara, Calif., USA), and thenhybridized overnight on Human Genome Arrays.

Alternatively, the protein products expressed from the mRNAs may beassayed by immunohistochemistry of tumour samples, solid phaseimmunoassay with microtitre plates, Western blotting, 2-dimensionalSDS-polyacrylamide gel electrophoresis, ELISA, flow cytometry and othermethods known in the art for detection of specific proteins. Detectionmethods would include the use of site specific antibodies. The skilledperson will recognize that all such well-known techniques for detectionof upregulation of IAP, detection of IAP variants or mutants, ordetection of 11q22 amplification could be applicable in the presentcase.

Abnormal levels of proteins such as IAP can be measured using standardprotein assays, for example, those assays described herein. Elevatedlevels or overexpression could also be detected in a tissue sample, forexample, a tumour tissue by measuring the protein levels with an assaysuch as that from Chemicon International. The protein of interest wouldbe immunoprecipitated from the sample lysate and its levels measured.

Alternative methods for the measurement of the over expression orelevation of IAPs including the isoforms thereof, include themeasurement of microvessel density. This can for example be measuredusing methods described by Orre and Rogers (Int J Cancer (1999), 84(2),101-8). Assay methods also include the use of markers.

Therefore all of these techniques could also be used to identify tumoursparticularly suitable for treatment with the compounds of the invention.

Therefore in a further aspect of the invention includes use of acompound according to the invention for the manufacture of a medicamentfor the treatment or prophylaxis of a disease state or condition in apatient who has been screened and has been determined as suffering from,or being at risk of suffering from, a disease or condition which wouldbe susceptible to treatment with a compound having affinity for IAP(i.e. an IAP antagonist).

A further embodiment provides a method of treating a patient having, orat risk of having a disease or condition described herein (e.g. cancer)comprising administering an effective amount of a compound of formula(I).

Another aspect of the invention includes a compound of the invention foruse in the prophylaxis or treatment of cancer in a patient selected froma sub-population possessing overexpression of one or more of the IAPfamily members (e.g. cIAP and/or XIAP).

Another aspect of the invention includes a compound of the invention foruse in the prophylaxis or treatment of cancer in a patient selected aspossessing a cytogenetic abherration that results in overexpression ofIAPs, for example, a patient selected as possessing the 11q22amplification.

MRI determination of vessel normalization (e.g. using MRI gradient echo,spin echo, and contrast enhancement to measure blood volume, relativevessel size, and vascular permeability) in combination with circulatingbiomarkers may also be used to identify for treatment with a compound ofthe invention.

Thus a further aspect of the invention is a method for the diagnosis andtreatment of a disease state or condition mediated by a IAP, whichmethod comprises (i) screening a patient to determine whether a diseaseor condition from which the patient is or may be suffering is one whichwould be susceptible to treatment with a compound having affinity forIAP; and (ii) where it is indicated that the disease or condition fromwhich the patient is thus susceptible, thereafter administering to thepatient a compound of formula (I) and sub-groups or examples thereof asdefined herein.

Pharmaceutical Formulations

While it is possible for the active compound to be administered alone,it is preferable to present it as a pharmaceutical composition (e.g.formulation). In one embodiment this is a sterile pharmaceuticalcomposition.

Thus, the present invention further provides pharmaceuticalcompositions, as defined above, and methods of making a pharmaceuticalcomposition comprising (e.g admixing) at least one compound of formula(I) (and sub-groups thereof as defined herein), together with one ormore pharmaceutically acceptable excipients and optionally othertherapeutic or prophylactic agents, as described herein.

The pharmaceutically acceptable excipient(s) can be selected from, forexample, carriers (e.g. a solid, liquid or semi-solid carrier),adjuvants, diluents, fillers or bulking agents, granulating agents,coating agents, release-controlling agents, binding agents,disintegrants, lubricating agents, preservatives, antioxidants,buffering agents, suspending agents, thickening agents, flavouringagents, sweeteners, taste masking agents, stabilisers or any otherexcipients conventionally used in pharmaceutical compositions. Examplesof excipients for various types of pharmaceutical compositions are setout in more detail below.

The term “pharmaceutically acceptable” as used herein pertains tocompounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of a subject (e.g. human) without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio. Each carrier,excipient, etc. must also be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation.

Pharmaceutical compositions containing compounds of the formula (I) canbe formulated in accordance with known techniques, see for example,Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton,Pa., USA.

The pharmaceutical compositions can be in any form suitable for oral,parenteral, topical, intranasal, intrabronchial, sublingual, ophthalmic,otic, rectal, intra-vaginal, or transdermal administration. Where thecompositions are intended for parenteral administration, they can beformulated for intravenous, intramuscular, intraperitoneal, subcutaneousadministration or for direct delivery into a target organ or tissue byinjection, infusion or other means of delivery. The delivery can be bybolus injection, short term infusion or longer term infusion and can bevia passive delivery or through the utilisation of a suitable infusionpump or syringe driver.

Pharmaceutical formulations adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions which maycontain anti-oxidants, buffers, bacteriostats, co-solvents, surfaceactive agents, organic solvent mixtures, cyclodextrin complexationagents, emulsifying agents (for forming and stabilizing emulsionformulations), liposome components for forming liposomes, gellablepolymers for forming polymeric gels, lyophilisation protectants andcombinations of agents for, inter alia, stabilising the activeingredient in a soluble form and rendering the formulation isotonic withthe blood of the intended recipient. Pharmaceutical formulations forparenteral administration may also take the form of aqueous andnon-aqueous sterile suspensions which may include suspending agents andthickening agents (R. G. Strickly, Solubilizing Excipients in oral andinjectable formulations, Pharmaceutical Research, Vol 21(2) 2004, p201-230).

The formulations may be presented in unit-dose or multi-dose containers,for example sealed ampoules, vials and prefilled syringes, and may bestored in a freeze-dried (lyophilised) condition requiring only theaddition of the sterile liquid carrier, for example water forinjections, immediately prior to use. In one embodiment, the formulationis provided as an active pharmaceutical ingredient in a bottle forsubsequent reconstitution using an appropriate diluent.

The pharmaceutical formulation can be prepared by lyophilising acompound of formula (I), or sub-groups thereof. Lyophilisation refers tothe procedure of freeze-drying a composition. Freeze-drying andlyophilisation are therefore used herein as synonyms.

Extemporaneous injection solutions and suspensions may be prepared fromsterile powders, granules and tablets.

Pharmaceutical compositions of the present invention for parenteralinjection can also comprise pharmaceutically acceptable sterile aqueousor non-aqueous solutions, dispersions, suspensions or emulsions as wellas sterile powders for reconstitution into sterile injectable solutionsor dispersions just prior to use.

Examples of suitable aqueous and nonaqueous carriers, diluents, solventsor vehicles include water, ethanol, polyols (such as glycerol, propyleneglycol, polyethylene glycol, and the like), carboxymethylcellulose andsuitable mixtures thereof, vegetable oils (such as sunflower oil,safflower oil, corn oil or olive oil), and injectable organic esterssuch as ethyl oleate. Proper fluidity can be maintained, for example, bythe use of thickening or coating materials such as lecithin, by themaintenance of the required particle size in the case of dispersions,and by the use of surfactants.

The compositions of the present invention may also contain adjuvantssuch as preservatives, wetting agents, emulsifying agents, anddispersing agents. Prevention of the action of microorganisms may beensured by the inclusion of various antibacterial and antifungal agents,for example, paraben, chlorobutanol, phenol, sorbic acid, and the like.It may also be desirable to include agents to adjust tonicity such assugars, sodium chloride, and the like. Prolonged absorption of theinjectable pharmaceutical form may be brought about by the inclusion ofagents which delay absorption such as aluminum monostearate and gelatin.

In one particular embodiment of the invention, the pharmaceuticalcomposition is in a form suitable for i.v. administration, for exampleby injection or infusion. For intravenous administration, the solutioncan be dosed as is, or can be injected into an infusion bag (containinga pharmaceutically acceptable excipient, such as 0.9% saline or 5%dextrose), before administration.

In another particular embodiment, the pharmaceutical composition is in aform suitable for sub-cutaneous (s.c.) administration.

Pharmaceutical dosage forms suitable for oral administration includetablets (coated or uncoated), capsules (hard or soft shell), caplets,pills, lozenges, syrups, solutions, powders, granules, elixirs andsuspensions, sublingual tablets, wafers or patches such as buccalpatches.

Thus, tablet compositions can contain a unit dosage of active compoundtogether with an inert diluent or carrier such as a sugar or sugaralcohol, eg; lactose, sucrose, sorbitol or mannitol; and/or a non-sugarderived diluent such as sodium carbonate, calcium phosphate, calciumcarbonate, or a cellulose or derivative thereof such as microcrystallinecellulose (MCC), methyl cellulose, ethyl cellulose, hydroxypropyl methylcellulose, and starches such as corn starch. Tablets may also containsuch standard ingredients as binding and granulating agents such aspolyvinylpyrrolidone, disintegrants (e.g. swellable crosslinked polymerssuch as crosslinked carboxymethylcellulose), lubricating agents (e.g.stearates), preservatives (e.g. parabens), antioxidants (e.g. BHT),buffering agents (for example phosphate or citrate buffers), andeffervescent agents such as citrate/bicarbonate mixtures. Suchexcipients are well known and do not need to be discussed in detailhere.

Tablets may be designed to release the drug either upon contact withstomach fluids (immediate release tablets) or to release in a controlledmanner (controlled release tablets) over a prolonged period of time orwith a specific region of the GI tract.

Capsule formulations may be of the hard gelatin or soft gelatin varietyand can contain the active component in solid, semi-solid, or liquidform. Gelatin capsules can be formed from animal gelatin or synthetic orplant derived equivalents thereof.

The solid dosage forms (eg; tablets, capsules etc.) can be coated orun-coated. Coatings may act either as a protective film (e.g. a polymer,wax or varnish) or as a mechanism for controlling drug release or foraesthetic or identification purposes. The coating (e.g. a Eudragit™ typepolymer) can be designed to release the active component at a desiredlocation within the gastro-intestinal tract. Thus, the coating can beselected so as to degrade under certain pH conditions within thegastrointestinal tract, thereby selectively release the compound in thestomach or in the ileum, duodenum, jejenum or colon.

Instead of, or in addition to, a coating, the drug can be presented in asolid matrix comprising a release controlling agent, for example arelease delaying agent which may be adapted to release the compound in acontrolled manner in the gastrointestinal tract. Alternatively the drugcan be presented in a polymer coating e.g. a polymethacrylate polymercoating, which may be adapted to selectively release the compound underconditions of varying acidity or alkalinity in the gastrointestinaltract. Alternatively, the matrix material or release retarding coatingcan take the form of an erodible polymer (e.g. a maleic anhydridepolymer) which is substantially continuously eroded as the dosage formpasses through the gastrointestinal tract. In another alternative, thecoating can be designed to disintegrate under microbial action in thegut. As a further alternative, the active compound can be formulated ina delivery system that provides osmotic control of the release of thecompound. Osmotic release and other delayed release or sustained releaseformulations (for example formulations based on ion exchange resins) maybe prepared in accordance with methods well known to those skilled inthe art.

The compound of formula (I) may be formulated with a carrier andadministered in the form of nanoparticles, the increased surface area ofthe nanoparticles assisting their absorption. In addition, nanoparticlesoffer the possibility of direct penetration into the cell. Nanoparticledrug delivery systems are described in “Nanoparticle Technology for DrugDelivery”, edited by Ram B Gupta and Uday B. Kompella, InformaHealthcare, ISBN 9781574448573, published 13th March 2006. Nanoparticlesfor drug delivery are also described in J. Control. Release, 2003, 91(1-2), 167-172, and in Sinha et al., Mol. Cancer Ther. August 1, (2006)5, 1909.

The pharmaceutical compositions typically comprise from approximately 1%(w/w) to approximately 95% (w/w) active ingredient and from 99% (w/w) to5% (w/w) of a pharmaceutically acceptable excipient or combination ofexcipients. Particularly, the compositions comprise from approximately20% (w/w) to approximately 90%,% (w/w) active ingredient and from 80%(w/w) to 10% of a pharmaceutically acceptable excipient or combinationof excipients. The pharmaceutical compositions comprise fromapproximately 1% to approximately 95%, particularly from approximately20% to approximately 90%, active ingredient. Pharmaceutical compositionsaccording to the invention may be, for example, in unit dose form, suchas in the form of ampoules, vials, suppositories, pre-filled syringes,dragées, tablets or capsules.

The pharmaceutically acceptable excipient(s) can be selected accordingto the desired physical form of the formulation and can, for example, beselected from diluents (e.g solid diluents such as fillers or bulkingagents; and liquid diluents such as solvents and co-solvents),disintegrants, buffering agents, lubricants, flow aids, releasecontrolling (e.g. release retarding or delaying polymers or waxes)agents, binders, granulating agents, pigments, plasticizers,antioxidants, preservatives, flavouring agents, taste masking agents,tonicity adjusting agents and coating agents.

The skilled person will have the expertise to select the appropriateamounts of ingredients for use in the formulations. For example tabletsand capsules typically contain 0-20% disintegrants, 0-5% lubricants,0-5% flow aids and/or 0-99% (w/w) fillers/or bulking agents (dependingon drug dose). They may also contain 0-10% (w/w) polymer binders, 0-5%(w/w) antioxidants, 0-5% (w/w) pigments. Slow release tablets would inaddition contain 0-99% (w/w) release-controlling (e.g. delaying)polymers (depending on dose). The film coats of the tablet or capsuletypically contain 0-10% (w/w) polymers, 0-3% (w/w) pigments, and/or 0-2%(w/w) plasticizers.

Parenteral formulations typically contain 0-20% (w/w) buffers, 0-50%(w/w) cosolvents, and/or 0-99% (w/w) Water for Injection (WFI)(depending on dose and if freeze dried). Formulations for intramusculardepots may also contain 0-99% (w/w) oils.

Pharmaceutical compositions for oral administration can be obtained bycombining the active ingredient with solid carriers, if desiredgranulating a resulting mixture, and processing the mixture, if desiredor necessary, after the addition of appropriate excipients, intotablets, dragee cores or capsules. It is also possible for them to beincorporated into a polymer or waxy matrix that allow the activeingredients to diffuse or be released in measured amounts.

The compounds of the invention can also be formulated as soliddispersions. Solid dispersions are homogeneous extremely fine dispersephases of two or more solids. Solid solutions (molecularly dispersesystems), one type of solid dispersion, are well known for use inpharmaceutical technology (see (Chiou and Riegelman, J. Pharm. Sci., 60,1281-1300 (1971)) and are useful in increasing dissolution rates andincreasing the bioavailability of poorly water-soluble drugs.

This invention also provides solid dosage forms comprising the solidsolution described above. Solid dosage forms include tablets, capsules,chewable tablets and dispersible or effervescent tablets. Knownexcipients can be blended with the solid solution to provide the desireddosage form. For example, a capsule can contain the solid solutionblended with (a) a disintegrant and a lubricant, or (b) a disintegrant,a lubricant and a surfactant. In addition a capsule can contain abulking agent, such as lactose or microcrystalline cellulose. A tabletcan contain the solid solution blended with at least one disintegrant, alubricant, a surfactant, a bulking agent and a glidant. A chewabletablet can contain the solid solution blended with a bulking agent, alubricant, and if desired an additional sweetening agent (such as anartificial sweetener), and suitable flavours. Solid solutions may alsobe formed by spraying solutions of drug and a suitable polymer onto thesurface of inert carriers such as sugar beads (‘non-pareils’). Thesebeads can subsequently be filled into capsules or compressed intotablets.

The pharmaceutical formulations may be presented to a patient in“patient packs” containing an entire course of treatment in a singlepackage, usually a blister pack. Patient packs have an advantage overtraditional prescriptions, where a pharmacist divides a patient's supplyof a pharmaceutical from a bulk supply, in that the patient always hasaccess to the package insert contained in the patient pack, normallymissing in patient prescriptions. The inclusion of a package insert hasbeen shown to improve patient compliance with the physician'sinstructions.

Compositions for topical use and nasal delivery include ointments,creams, sprays, patches, gels, liquid drops and inserts (for exampleintraocular inserts). Such compositions can be formulated in accordancewith known methods.

Examples of formulations for rectal or intra-vaginal administrationinclude pessaries and suppositories which may be, for example, formedfrom a shaped moldable or waxy material containing the active compound.Solutions of the active compound may also be used for rectaladministration.

Compositions for administration by inhalation may take the form ofinhalable powder compositions or liquid or powder sprays, and can beadministrated in standard form using powder inhaler devices or aerosoldispensing devices. Such devices are well known. For administration byinhalation, the powdered formulations typically comprise the activecompound together with an inert solid powdered diluent such as lactose.

The compounds of the formula (I) will generally be presented in unitdosage form and, as such, will typically contain sufficient compound toprovide a desired level of biological activity. For example, aformulation may contain from 1 nanogram to 2 grams of active ingredient,e.g. from 1 nanogram to 2 milligrams of active ingredient. Within theseranges, particular sub-ranges of compound are 0.1 milligrams to 2 gramsof active ingredient (more usually from 10 milligrams to 1 gram, e.g. 50milligrams to 500 milligrams), or 1 microgram to 20 milligrams (forexample 1 microgram to 10 milligrams, e.g. 0.1 milligrams to 2milligrams of active ingredient).

For oral compositions, a unit dosage form may contain from 1 milligramto 2 grams, more typically 10 milligrams to 1 gram, for example 50milligrams to 1 gram, e.g. 100 miligrams to 1 gram, of active compound.

The active compound will be administered to a patient in need thereof(for example a human or animal patient) in an amount sufficient toachieve the desired therapeutic effect.

Methods of Treatment

The compounds of the formula (I) and sub-groups as defined herein may beuseful in the prophylaxis or treatment of a range of disease states orconditions mediated by IAP. Thus, according to a further aspect of theinvention there is provided a method of treating a disease state orcondition mediated by IAP, such as an XIAP and/or cIAP (e.g. cancer)which comprises administering to a subject in need thereof a compound offormula (I) as described herein. According to a further aspect of theinvention there is provided a method of treating a disease state orcondition (e.g. cancer) which overexpresses IAP, such as an XIAP and/orcIAP which comprises administering to a subject in need thereof acompound of formula (I) as described herein. Examples of such diseasestates and conditions are set out above, and in particular includecancer.

The compounds are generally administered to a subject in need of suchadministration, for example a human or animal patient, particularly ahuman.

The compounds will typically be administered in amounts that aretherapeutically or prophylactically useful and which generally arenon-toxic. However, in certain situations (for example in the case oflife threatening diseases), the benefits of administering a compound ofthe formula (I) may outweigh the disadvantages of any toxic effects orside effects, in which case it may be considered desirable to administercompounds in amounts that are associated with a degree of toxicity.

The compounds may be administered over a prolonged term to maintainbeneficial therapeutic effects or may be administered for a short periodonly. Alternatively they may be administered in a continuous manner orin a manner that provides intermittent dosing (e.g. a pulsatile manner).

A typical daily dose of the compound of formula (I) can be in the rangefrom 100 picograms to 100 milligrams per kilogram of body weight, moretypically 5 nanograms to 25 milligrams per kilogram of bodyweight, andmore usually 10 nanograms to 15 milligrams per kilogram (e.g. 10nanograms to 10 milligrams, and more typically 1 microgram per kilogramto 20 milligrams per kilogram, for example 1 microgram to 10 milligramsper kilogram) per kilogram of bodyweight although higher or lower dosesmay be administered where required. The compound of the formula (I) canbe administered on a daily basis or on a repeat basis every 2, or 3, or4, or 5, or 6, or 7, or 10 or 14, or 21, or 28 days for example.

The compounds of the invention may be administered orally in a range ofdoses, for example 1 to 1500 mg, 2 to 800 mg, or 5 to 500 mg, e.g. 2 to200 mg or 10 to 1000 mg, particular examples of doses including 10, 20,50 and 80 mg. The compound may be administered once or more than onceeach day. The compound can be administered continuously (i.e. takenevery day without a break for the duration of the treatment regimen).Alternatively, the compound can be administered intermittently (i.e.taken continuously for a given period such as a week, then discontinuedfor a period such as a week and then taken continuously for anotherperiod such as a week and so on throughout the duration of the treatmentregimen). Examples of treatment regimens involving intermittentadministration include regimens wherein administration is in cycles ofone week on, one week off; or two weeks on, one week off; or three weekson, one week off; or two weeks on, two weeks off; or four weeks on twoweeks off; or one week on three weeks off—for one or more cycles, e.g.2, 3, 4, 5, 6, 7, 8, 9 or 10 or more cycles.

In one particular dosing schedule, a patient will be given an infusionof a compound of the formula (I) for periods of one hour daily for up toten days in particular up to five days for one week, and the treatmentrepeated at a desired interval such as two to four weeks, in particularevery three weeks.

More particularly, a patient may be given an infusion of a compound ofthe formula (I) for periods of one hour daily for 5 days and thetreatment repeated every three weeks.

In another particular dosing schedule, a patient is given an infusionover 30 minutes to 1 hour followed by maintenance infusions of variableduration, for example 1 to 5 hours, e.g. 3 hours.

In a further particular dosing schedule, a patient is given a continuousinfusion for a period of 12 hours to 5 days, an in particular acontinuous infusion of 24 hours to 72 hours.

In another particular dosing schedule, a patient is given the compoundorally once a week.

In another particular dosing schedule, a patient is given the compoundorally once-daily for between 7 and 28 days such as 7, 14 or 28 days.

In another particular dosing schedule, a patient is given the compoundorally once-daily for 1 day, 2 days, 3 days, 5 days or 1 week followedby the required amount of days off to complete a one or two week cycle.

In another particular dosing schedule, a patient is given the compoundorally once-daily for 2 weeks followed by 2 weeks off.

In another particular dosing schedule, a patient is given the compoundorally once-daily for 2 weeks followed by 1 week off.

In another particular dosing schedule, a patient is given the compoundorally once-daily for 1 week followed by 1 week off.

Ultimately, however, the quantity of compound administered and the typeof composition used will be commensurate with the nature of the diseaseor physiological condition being treated and will be at the discretionof the physician.

It has been discovered that IAP antagonists can be used as a singleagent or in combination with other anticancer agents. For example, itmay be beneficial to combine an antagonist that induces apoptosis withanother agent which acts via a different mechanism to regulate cellgrowth thus treating two of the characteristic features of cancerdevelopment. Combination experiments can be performed, for example, asdescribed in Chou T C, Talalay P. Quantitative analysis of dose-effectrelationships: the combined effects of multiple drugs or enzymeinhibitors. Adv Enzyme Regulat 1984; 22: 27-55.

The compounds as defined herein can be administered as the soletherapeutic agent or they can be administered in combination therapywith one of more other compounds (or therapies) for treatment of aparticular disease state, for example a neoplastic disease such as acancer as hereinbefore defined. For the treatment of the aboveconditions, the compounds of the invention may be advantageouslyemployed in combination with one or more other medicinal agents, moreparticularly, with other anti-cancer agents or adjuvants (supportingagents in the therapy) in cancer therapy. Examples of other therapeuticagents or treatments that may be administered together (whetherconcurrently or at different time intervals) with the compounds of theformula (I) include but are not limited to:

-   -   Topoisomerase I inhibitors;    -   Antimetabolites;    -   Tubulin targeting agents;    -   DNA binder and topoisomerase II inhibitors;    -   Alkylating Agents;    -   Monoclonal Antibodies;    -   Anti-Hormones;    -   Signal Transduction Inhibitors;    -   Proteasome Inhibitors;    -   DNA methyl transferase inhibitors;    -   Cytokines and retinoids;    -   Chromatin targeted therapies;    -   Radiotherapy; and    -   Other therapeutic or prophylactic agents.

Particular examples of anti-cancer agents or adjuvants (or saltsthereof), include but are not limited to any of the agents selected fromgroups (i)-(xlvi), and optionally group (xlvii), below:

-   (i) Platinum compounds, for example cisplatin (optionally combined    with amifostine), carboplatin or oxaliplatin;-   (ii) Taxane compounds, for example paclitaxel, paclitaxel protein    bound particles (Abraxane™), docetaxel, cabazitaxel or larotaxel;-   (iii) Topoisomerase I inhibitors, for example camptothecin    compounds, for example camptothecin, irinotecan(CPT11), SN-38, or    topotecan;-   (iv) Topoisomerase II inhibitors, for example anti-tumour    epipodophyllotoxins or podophyllotoxin derivatives for example    etoposide, or teniposide;-   (v) Vinca alkaloids, for example vinblastine, vincristine, liposomal    vincristine (Onco-TCS), vinorelbine, vindesine, vinflunine or    vinvesir;-   (vi) Nucleoside derivatives, for example 5-fluorouracil (5-FU,    optionally in combination with leucovorin), gemcitabine,    capecitabine, tegafur, UFT, S1, cladribine, cytarabine (Ara-C,    cytosine arabinoside), fludarabine, clofarabine, or nelarabine;-   (vii) Antimetabolites, for example clofarabine, aminopterin, or    methotrexate, azacitidine, cytarabine, floxuridine, pentostatin,    thioguanine, thiopurine, 6-mercaptopurine, or hydroxyurea    (hydroxycarbamide);-   (viii) Alkylating agents, such as nitrogen mustards or nitrosourea,    for example cyclophosphamide, chlorambucil, carmustine (BCNU),    bendamustine, thiotepa, melphalan, treosulfan, lomustine (CCNU),    altretamine, busulfan, dacarbazine, estramustine, fotemustine,    ifosfamide (optionally in combination with mesna), pipobroman,    procarbazine, streptozocin, temozolomide, uracil, mechlorethamine,    methylcyclohexylchloroethylnitrosurea, or nimustine (ACNU);-   (ix) Anthracyclines, anthracenediones and related drugs, for example    daunorubicin, doxorubicin (optionally in combination with    dexrazoxane), liposomal formulations of doxorubicin (eg. Caelyx™,    Myocet™, Doxil™), idarubicin, mitoxantrone, epirubicin, amsacrine,    or valrubicin;-   (x) Epothilones, for example ixabepilone, patupilone, BMS-310705,    KOS-862 and ZK-EPO, epothilone A, epothilone B, desoxyepothilone B    (also known as epothilone D or KOS-862), aza-epothilone B (also    known as BMS-247550), aulimalide, isolaulimalide, or    luetherobin; (xi) DNA methyl transferase inhibitors, for example    temozolomide, azacytidine or decitabine, or SGI-110;-   (xii) Antifolates, for example methotrexate, pemetrexed disodium, or    raltitrexed;-   (xiii) Cytotoxic antibiotics, for example antinomycin D, bleomycin,    mitomycin C, dactinomycin, carminomycin, daunomycin, levamisole,    plicamycin, or mithramycin;-   (xiv) Tubulin-binding agents, for example combrestatin, colchicines    or nocodazole;-   (xv) Signal Transduction inhibitors such as Kinase inhibitors (e.g.    EGFR (epithelial growth factor receptor) inhibitors, VEGFR (vascular    endothelial growth factor receptor) inhibitors, PDGFR    (platelet-derived growth factor receptor) inhibitors, MTKI (multi    target kinase inhibitors), Raf inhibitors, mTOR inhibitors for    example imatinib mesylate, erlotinib, gefitinib, dasatinib,    lapatinib, dovotinib, axitinib, nilotinib, vandetanib, vatalinib,    pazopanib, sorafenib, sunitinib, temsirolimus, everolimus (RAD 001),    vemurafenib (PLX4032/RG7204), dabrafenib, encorafenib or an IκB    kinase inhibitor such as SAR-113945, bardoxolone, BMS-066,    BMS-345541, IMD-0354, IMD-2560, or IMD-1041, or MEK inhibitors such    as Selumetinib (AZD6244) and Trametinib (GSK121120212);-   (xvi) Aurora kinase inhibitors for example AT9283, barasertib    (AZD1152), TAK-901, MK0457 (VX680), cenisertib (R-763), danusertib    (PHA-739358), alisertib (MLN-8237), or MP-470;-   (xvii) CDK inhibitors for example AT7519, roscovitine, seliciclib,    alvocidib (flavopiridol), dinaciclib (SCH-727965),    7-hydroxy-staurosporine (UCN-01), JNJ-7706621, BMS-387032 (a.k.a.    SNS-032), PHA533533, PD332991, ZK-304709, or AZD-5438;-   (xviii) PKA/B inhibitors and PKB (akt) pathway inhibitors for    example AKT inhibitors such as KRX-0401 (perifosine/NSC 639966),    ipatasertib (GDC-0068; RG-7440), afuresertib (GSK-2110183; 2110183),    MK-2206, MK-8156, AT13148, AZD-5363, triciribine phosphate (VQD-002;    triciribine phosphate monohydrate (API-2; TCN-P; TCN-PM; VD-0002),    RX-0201, NL-71-101, SR-13668, PX-316, AT13148, AZ-5363, Semaphore,    SF1126, or Enzastaurin HCl (LY317615) or MTOR inhibitors such as    rapamycin analogues such as RAD 001 (everolimus), CCI 779    (temsirolemus), AP23573 and ridaforolimus, sirolimus (originally    known as rapamycin), AP23841 and AP23573, calmodulin inhibitors e.g.    CBP-501 (forkhead translocation inhibitors), enzastaurin HCl    (LY317615) or PI3K Inhibitors such as dactolisib (BEZ235),    buparlisib (BKM-120; NVP-BKM-120), BYL719, copanlisib (BAY-80-6946),    ZSTK-474, CUDC-907, apitolisib (GDC-0980; RG-7422), pictilisib    (pictrelisib, GDC-0941, RG-7321), GDC-0032, GDC-0068, GSK-2636771,    idelalisib (formerly CAL-101, GS 1101, GS-1101), MLN1117 (INK1117),    MLN0128 (INK128), IPI-145 (INK1197), LY-3023414, ipatasertib,    afuresertib, MK-2206, MK-8156, LY-3023414, LY294002, SF1126 or    PI-103, or sonolisib (PX-866);-   (xix) Hsp90 inhibitors for example AT13387, herbimycin, geldanamycin    (GA), 17-allylamino-17-desmethoxygeldanamycin (17-AAG) e.g.    NSC-330507, Kos-953 and CNF-1010,    17-dimethylaminoethylamino-17-demethoxygeldanamycin hydrochloride    (17-DMAG) e.g. NSC-707545 and Kos-1022, NVP-AUY922 (VER-52296),    NVP-BEP800, CNF-2024 (BUB-021 an oral purine), ganetespib    (STA-9090), SNX-5422 (SC-102112) or IPI-504;-   (xx) Monoclonal Antibodies (unconjugated or conjugated to    radioisotopes, toxins or other agents), antibody derivatives and    related agents, such as anti-CD, anti-VEGFR, anti-HER2, anti-CTLA4,    anti-PD-1 or anti-EGFR antibodies, for example rituximab (CD20),    ofatumumab (CD20), ibritumomab tiuxetan (CD20), GA101 (CD20),    tositumomab (CD20), epratuzumab (CD22), lintuzumab (CD33),    gemtuzumab ozogamicin (CD33), alemtuzumab (CD52), galiximab (CD80),    trastuzumab (HER2 antibody), pertuzumab (HER2), trastuzumab-DM1    (HER2), ertumaxomab (HER2 and CD3), cetuximab (EGFR), panitumumab    (EGFR), necitumumab (EGFR), nimotuzumab (EGFR), bevacizumab (VEGF),    catumaxumab (EpCAM and CD3), abagovomab (CA125), farletuzumab    (folate receptor), elotuzumab (CS1), denosumab (RANK ligand),    figitumumab (IGF1R), CP751,871 (IGF1R), mapatumumab (TRAIL    receptor), metMAB (met), mitumomab (GD3 ganglioside), naptumomab    estafenatox (5T4), siltuximab (IL6), or immunomodulating agents such    as CTLA-4 blocking antibodies and/or antibodies against PD-1 and    PD-L1 and/or PD-L2 for example ipilimumab (CTLA4), MK-3475    (pembrolizumab, formerly lambrolizumab, anti-PD-1), nivolumab    (anti-PD-1), BMS-936559 (anti-PD-L1), MPDL320A, AMP-514 or MED14736    (anti-PD-L1), or tremelimumab (formerly ticilimumab, CP-675,206,    anti-CTLA-4);-   (xxi) Estrogen receptor antagonists or selective estrogen receptor    modulators (SERMs) or inhibitors of estrogen synthesis, for example    tamoxifen, fulvestrant, toremifene, droloxifene, faslodex, or    raloxifene;-   (xxii) Aromatase inhibitors and related drugs, such as exemestane,    anastrozole, letrazole, testolactone aminoglutethimide, mitotane or    vorozole;-   (xxiii) Antiandrogens (i.e. androgen receptor antagonists) and    related agents for example bicalutamide, nilutamide, flutamide,    cyproterone, or ketoconazole;-   (xxiv) Hormones and analogues thereof such as medroxyprogesterone,    diethylstilbestrol (a.k.a. diethylstilboestrol) or octreotide;-   (xxv) Steroids for example dromostanolone propionate, megestrol    acetate, nandrolone (decanoate, phenpropionate), fluoxymestrone or    gossypol,-   (xxvii) Steroidal cytochrome P450 17alpha-hydroxylase-17,20-lyase    inhibitor (CYP17), e.g. abiraterone;-   (xxvii) Gonadotropin releasing hormone agonists or antagonists    (GnRAs) for example abarelix, goserelin acetate, histrelin acetate,    leuprolide acetate, triptorelin, buserelin, or deslorelin;-   (xxviii) Glucocorticoids, for example prednisone, prednisolone,    dexamethasone;-   (xxix) Differentiating agents, such as retinoids, rexinoids, vitamin    D or retinoic acid and retinoic acid metabolism blocking agents (RAM    BA) for example accutane, alitretinoin, bexarotene, or tretinoin;-   (xxx) Farnesyltransferase inhibitors for example tipifarnib;-   (xxxi) Chromatin targeted therapies such as histone deacetylase    (HDAC) inhibitors for example panobinostat, resminostat,    abexinostat, vorinostat, romidepsin, belinostat, entinostat,    quisinostat, pracinostat, tefinostat, mocetinostat, givinostat,    CUDC-907, CUDC-101, ACY-1215, MGCD-290, EVP-0334, RG-2833, 4SC-202,    romidepsin, AR-42 (Ohio State University), CG-200745, valproic acid,    CKD-581, sodium butyrate, suberoylanilide hydroxamide acid (SAHA),    depsipeptide (FR 901228), dacinostat (NVP-LAQ824),    R306465/JNJ-16241199, JNJ-26481585, trichostatin A, chlamydocin,    A-173, JNJ-MGCD-0103, PXD-101, or apicidin;-   (xxxii) Proteasome Inhibitors for example bortezomib, carfilzomib,    delanzomib (CEP-18770), ixazomib (MLN-9708), oprozomib (ONX-0912) or    marizomib;-   (xxxiii) Photodynamic drugs for example porfimer sodium or    temoporfin;-   (xxxiv) Marine organism-derived anticancer agents such as    trabectidin;-   (xxxv) Radiolabelled drugs for radioimmunotherapy for example with a    beta particle-emitting isotope (e.g., Iodine-131, Yittrium-90) or an    alpha particle-emitting isotope (e.g., Bismuth-213 or Actinium-225)    for example ibritumomab or Iodine tositumomab;-   (xxxvi) Telomerase inhibitors for example telomestatin;-   (xxxvii) Matrix metalloproteinase inhibitors for example batimastat,    marimastat, prinostat or metastat;-   (xxxviii) Recombinant interferons (such as interferon-γ and    interferon α) and interleukins (e.g. interleukin 2), for example    aldesleukin, denileukin diftitox, interferon alfa 2a, interferon    alfa 2b, or peginterferon alfa 2b;-   (xxxix) Selective immunoresponse modulators for example thalidomide,    or lenalidomide;-   (xl) Therapeutic Vaccines such as sipuleucel-T (Provenge) or    OncoVex;-   (xli) Cytokine-activating agents include Picibanil, Romurtide,    Sizofiran, Virulizin, or Thymosin;-   (xlii) Arsenic trioxide;-   (xliii) Inhibitors of G-protein coupled receptors (GPCR) for example    atrasentan;-   (xliv) Enzymes such as L-asparaginase, pegaspargase, rasburicase, or    pegademase;-   (xlv) DNA repair inhibitors such as PARP inhibitors for example,    olaparib, velaparib, iniparib, INO-1001, AG-014699, or ONO-2231;-   (xlvi) Agonists of Death receptor (e.g. TNF-related apoptosis    inducing ligand (TRAIL) receptor), such as mapatumumab (formerly    HGS-ETR1), conatumumab (formerly AMG 655), PRO95780, lexatumumab,    dulanermin, CS-1008, apomab or recombinant TRAIL ligands such as    recombinant Human TRAILL/Apo2 Ligand;-   (xlvii) Prophylactic agents (adjuncts); i.e. agents that reduce or    alleviate some of the side effects associated with chemotherapy    agents, for example    -   anti-emetic agents,    -   agents that prevent or decrease the duration of        chemotherapy-associated neutropenia and prevent complications        that arise from reduced levels of platelets, red blood cells or        white blood cells, for example interleukin-11 (e.g. oprelvekin),        erythropoietin (EPO) and analogues thereof (e.g. darbepoetin        alfa), colony-stimulating factor analogs such as granulocyte        macrophage-colony stimulating factor (GM-CSF) (e.g.        sargramostim), and granulocyte-colony stimulating factor (G-CSF)        and analogues thereof (e.g. filgrastim, pegfilgrastim),    -   agents that inhibit bone resorption such as denosumab or        bisphosphonates e.g. zoledronate, zoledronic acid, pamidronate        and ibandronate,    -   agents that suppress inflammatory responses such as        dexamethasone, prednisone, and prednisolone,    -   agents used to reduce blood levels of growth hormone and IGF-I        (and other hormones) in patients with acromegaly or other rare        hormone-producing tumours, such as synthetic forms of the        hormone somatostatin e.g. octreotide acetate,    -   antidote to drugs that decrease levels of folic acid such as        leucovorin, or folinic acid,    -   agents for pain e.g. opiates such as morphine, diamorphine and        fentanyl,    -   non-steroidal anti-inflammatory drugs (NSAID) such as COX-2        inhibitors for example celecoxib, etoricoxib and lumiracoxib,    -   agents for mucositis e.g. palifermin,    -   agents for the treatment of side-effects including anorexia,        cachexia, oedema or thromoembolic episodes, such as megestrol        acetate.

In one embodiment the anticancer is selected from recombinantinterferons (such as interferon-γ and interferon α) and interleukins(e.g. interleukin 2), for example aldesleukin, denileukin diftitox,interferon alfa 2a, interferon alfa 2b, or peginterferon alfa 2b;interferon-α2 (500 μ/ml) in particular interferon-β; and signaltransduction inhibitors such as kinase inhibitors (e.g. EGFR (epithelialgrowth factor receptor) inhibitors, VEGFR (vascular endothelial growthfactor receptor) inhibitors, PDGFR (platelet-derived growth factorreceptor) inhibitors, MTKI (multi target kinase inhibitors), Rafinhibitors, mTOR inhibitors for example imatinib mesylate, erlotinib,gefitinib, dasatinib, lapatinib, dovotinib, axitinib, nilotinib,vandetanib, vatalinib, pazopanib, sorafenib, sunitinib, temsirolimus,everolimus (RAD 001), vemurafenib (PLX4032/RG7204), dabrafenib,encorafenib or an IκB kinase inhibitor such as SAR-113945, bardoxolone,BMS-066, BMS-345541, IMD-0354, IMD-2560, or IMD-1041, or MEK inhibitorssuch as Selumetinib (AZD6244) and Trametinib (GSK121120212), inparticular Raf inhibitors (e.g. vemurafenib) or MEK inhibitors (e.g.trametinib).

Each of the compounds present in the combinations of the invention maybe given in individually varying dose schedules and via differentroutes. As such, the posology of each of the two or more agents maydiffer: each may be administered at the same time or at different times.A person skilled in the art would know through his or her common generalknowledge the dosing regimes and combination therapies to use. Forexample, the compound of the invention may be using in combination withone or more other agents which are administered according to theirexisting combination regimen. Examples of standard combination regimensare provided below.

The taxane compound is advantageously administered in a dosage of 50 to400 mg per square meter (mg/m²) of body surface area, for example 75 to250 mg/m², particularly for paclitaxel in a dosage of about 175 to 250mg/m² and for docetaxel in about 75 to 150 mg/m² per course oftreatment.

The camptothecin compound is advantageously administered in a dosage of0.1 to 400 mg per square meter (mg/m²) of body surface area, for example1 to 300 mg/m², particularly for irinotecan in a dosage of about 100 to350 mg/m² and for topotecan in about 1 to 2 mg/m² per course oftreatment.

The anti-tumour podophyllotoxin derivative is advantageouslyadministered in a dosage of 30 to 300 mg per square meter (mg/m²) ofbody surface area, for example 50 to 250 mg/m², particularly foretoposide in a dosage of about 35 to 100 mg/m² and for teniposide inabout 50 to 250 mg/m² per course of treatment.

The anti-tumour vinca alkaloid is advantageously administered in adosage of 2 to 30 mg per square meter (mg/m²) of body surface area,particularly for vinblastine in a dosage of about 3 to 12 mg/m², forvincristine in a dosage of about 1 to 2 mg/m², and for vinorelbine indosage of about 10 to 30 mg/m² per course of treatment.

The anti-tumour nucleoside derivative is advantageously administered ina dosage of 200 to 2500 mg per square meter (mg/m²) of body surfacearea, for example 700 to 1500 mg/m², particularly for 5-FU in a dosageof 200 to 500 mg/m², for gemcitabine in a dosage of about 800 to 1200mg/m² and for capecitabine in about 1000 to 2500 mg/m² per course oftreatment.

The alkylating agents such as nitrogen mustard or nitrosourea isadvantageously administered in a dosage of 100 to 500 mg per squaremeter (mg/m²) of body surface area, for example 120 to 200 mg/m²,particularly for cyclophosphamide in a dosage of about 100 to 500 mg/m²,for chlorambucil in a dosage of about 0.1 to 0.2 mg/kg, for carmustinein a dosage of about 150 to 200 mg/m², and for lomustine in a dosage ofabout 100 to 150 mg/m² per course of treatment.

The anti-tumour anthracycline derivative is advantageously administeredin a dosage of 10 to 75 mg per square meter (mg/m²) of body surfacearea, for example 15 to 60 mg/m², particularly for doxorubicin in adosage of about 40 to 75 mg/m², for daunorubicin in a dosage of about 25to 45 mg/m², and for idarubicin in a dosage of about 10 to 15 mg/m² percourse of treatment.

The antiestrogen agent is advantageously administered in a dosage ofabout 1 to 100 mg daily depending on the particular agent and thecondition being treated. Tamoxifen is advantageously administered orallyin a dosage of 5 to 50 mg, particularly 10 to 20 mg twice a day,continuing the therapy for sufficient time to achieve and maintain atherapeutic effect. Toremifene is advantageously administered orally ina dosage of about 60 mg once a day, continuing the therapy forsufficient time to achieve and maintain a therapeutic effect.Anastrozole is advantageously administered orally in a dosage of about 1mg once a day. Droloxifene is advantageously administered orally in adosage of about 20-100 mg once a day. Raloxifene is advantageouslyadministered orally in a dosage of about 60 mg once a day. Exemestane isadvantageously administered orally in a dosage of about 25 mg once aday.

Antibodies are advantageously administered in a dosage of about 1 to 5mg per square meter (mg/m²) of body surface area, or as known in theart, if different. Trastuzumab is advantageously administered in adosage of 1 to 5 mg per square meter (mg/m²) of body surface area,particularly 2 to 4 mg/m² per course of treatment.

Where the compound of the formula (I) is administered in combinationtherapy with one, two, three, four or more other therapeutic agents(particularly one or two, more particularly one), the compounds can beadministered simultaneously or sequentially. In the latter case, the twoor more compounds will be administered within a period and in an amountand manner that is sufficient to ensure that an advantageous orsynergistic effect is achieved. When administered sequentially, they canbe administered at closely spaced intervals (for example over a periodof 5-10 minutes) or at longer intervals (for example 1, 2, 3, 4 or morehours apart, or even longer periods apart where required), the precisedosage regimen being commensurate with the properties of the therapeuticagent(s). These dosages may be administered for example once, twice ormore per course of treatment, which may be repeated for example every 7,14, 21 or 28 days.

In one embodiment is provided a compound of formula (I) for themanufacture of a medicament for use in therapy wherein said compound isused in combination with one, two, three, or four other therapeuticagents. In another embodiment is provided a medicament for treatingcancer which comprises a compound of formula (I) wherein said medicamentis used in combination with one, two, three, or four other therapeuticagents. The invention further provides use of a compound of formula (I)for the manufacture of a medicament for enhancing or potentiating theresponse rate in a patient suffering from a cancer where the patient isbeing treated with one, two, three, or four other therapeutic agents.

It will be appreciated that the particular method and order ofadministration and the respective dosage amounts and regimes for eachcomponent of the combination will depend on the particular othermedicinal agent and compound of the present invention beingadministered, their route of administration, the particular tumour beingtreated and the particular host being treated. The optimum method andorder of administration and the dosage amounts and regime can be readilydetermined by those skilled in the art using conventional methods and inview of the information set out herein.

The weight ratio of the compound according to the present invention andthe one or more other anticancer agent(s) when given as a combinationmay be determined by the person skilled in the art. Said ratio and theexact dosage and frequency of administration depends on the particularcompound according to the invention and the other anticancer agent(s)used, the particular condition being treated, the severity of thecondition being treated, the age, weight, gender, diet, time ofadministration and general physical condition of the particular patient,the mode of administration as well as other medication the individualmay be taking, as is well known to those skilled in the art.Furthermore, it is evident that the effective daily amount may belowered or increased depending on the response of the treated subjectand/or depending on the evaluation of the physician prescribing thecompounds of the instant invention. A particular weight ratio for thepresent compound of formula (I) and another anticancer agent may rangefrom 1/10 to 10/1, more in particular from 1/5 to 5/1, even more inparticular from 1/3 to 3/1.

The compounds of the invention may also be administered in conjunctionwith non-chemotherapeutic treatments such as radiotherapy, photodynamictherapy, gene therapy; surgery and controlled diets.

The compounds of the present invention also have therapeuticapplications in sensitising tumour cells for radiotherapy andchemotherapy. Hence the compounds of the present invention can be usedas “radiosensitizer” and/or “chemosensitizer” or can be given incombination with another “radiosensitizer” and/or “chemosensitizer”. Inone embodiment the compound of the invention is for use aschemosensitiser.

The term “radiosensitizer” is defined as a molecule administered topatients in therapeutically effective amounts to increase thesensitivity of the cells to ionizing radiation and/or to promote thetreatment of diseases which are treatable with ionizing radiation.

The term “chemosensitizer” is defined as a molecule administered topatients in therapeutically effective amounts to increase thesensitivity of cells to chemotherapy and/or promote the treatment ofdiseases which are treatable with chemotherapeutics.

In one embodiment the compound of the invention is administered with a“radiosensitizer” and/or “chemosensitizer”. In one embodiment thecompound of the invention is administered with an “immune sensitizer”.

The term “immune sensitizer” is defined as a molecule administered topatients in therapeutically effective amounts to increase thesensitivity of cells to an IAP antagonist for example, by promoting orincreasing the immune response for example by triggering release of TNF.

Many cancer treatment protocols currently employ radiosensitizers inconjunction with radiation of x-rays. Examples of x-ray activatedradiosensitizers include, but are not limited to, the following:metronidazole, misonidazole, desmethylmisonidazole, pimonidazole,etanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233, E09, RB 6145,nicotinamide, 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (lUdR),bromodeoxycytidine, fluorodeoxyuridine (FudR), hydroxyurea, cisplatin,and therapeutically effective analogs and derivatives of the same.

Photodynamic therapy (PDT) of cancers employs visible light as theradiation activator of the sensitizing agent. Examples of photodynamicradiosensitizers include the following, but are not limited to:hematoporphyrin derivatives, Photofrin, benzoporphyrin derivatives, tinetioporphyrin, pheoborbide-a, bacteriochlorophyll-a, naphthalocyanines,phthalocyanines, zinc phthalocyanine, and therapeutically effectiveanalogs and derivatives of the same.

Radiosensitizers may be administered in conjunction with atherapeutically effective amount of one or more other compounds,including but not limited to: compounds of the invention; compoundswhich promote the incorporation of radiosensitizers to the target cells;compounds which control the flow of therapeutics, nutrients, and/oroxygen to the target cells; chemotherapeutic agents which act on thetumour with or without additional radiation; or other therapeuticallyeffective compounds for treating cancer or other diseases.

Chemosensitizers may be administered in conjunction with atherapeutically effective amount of one or more other compounds,including but not limited to: compounds of the invention; compoundswhich promote the incorporation of chemosensitizers to the target cells;compounds which control the flow of therapeutics, nutrients, and/oroxygen to the target cells; chemotherapeutic agents which act on thetumour or other therapeutically effective compounds for treating canceror other disease. Calcium antagonists, for example verapamil, are founduseful in combination with antineoplastic agents to establishchemosensitivity in tumor cells resistant to accepted chemotherapeuticagents and to potentiate the efficacy of such compounds indrug-sensitive malignancies.

Examples of immune sensitizers include the following, but are notlimited to: immunomodulating agents, for example monoclonal antibodiessuch as immune checkpoint antibodies [e.g. CTLA-4 blocking antibodiesand/or antibodies against PD-1 and PD-L1 and/or PD-L2 for exampleipilimumab (CTLA4), MK-3475 (pembrolizumab, formerly lambrolizumab,anti-PD-1), nivolumab (anti-PD-1), BMS-936559 (anti-PD-L1), MPDL320A,AMP-514 or MED14736 (anti-PD-L1), or tremelimumab (formerly ticilimumab,CP-675,206, anti-CTLA-4)]; or Signal Transduction inhibitors; orcytokines (such as recombinant interferons); or oncolytic viruses; orimmune adjuvants (e.g. BCG).

Immune sensitizers may be administered in conjunction with atherapeutically effective amount of one or more other compounds,including but not limited to: compounds of the invention; compoundswhich promote the incorporation of immune sensitizers to the targetcells; compounds which control the flow of therapeutics, nutrients,and/or oxygen to the target cells; therapeutic agents which act on thetumour or other therapeutically effective compounds for treating canceror other disease.

For use in combination therapy with another chemotherapeutic agent, thecompound of the formula (I) and one, two, three, four or more othertherapeutic agents can be, for example, formulated together in a dosageform containing two, three, four or more therapeutic agents i.e. in aunitary pharmaceutical composition containing all agents. In analternative embodiment, the individual therapeutic agents may beformulated separately and presented together in the form of a kit,optionally with instructions for their use.

In one embodiment is provided a combination of a compound of formula (I)with one or more (e.g. 1 or 2) other therapeutic agents (e.g. anticanceragents as described above). In a further embodiment is provided acombination of an IAP antagonist as described herein and a PI3K/AKTpathway inhibitor selected from: apitolisib, buparlisib, Copanlisib,pictilisib, ZSTK-474, CUDC-907, GSK-2636771, LY-3023414, ipatasertib,afuresertib, MK-2206, MK-8156, Idelalisib, BEZ235 (dactolisib), BYL719,GDC-0980, GDC-0941, GDC-0032 and GDC-0068.

In another embodiment is provided a compound of formula (I) incombination with one or more (e.g. 1 or 2) other therapeutic agents(e.g. anticancer agents) for use in therapy, such as in the prophylaxisor treatment of cancer.

In one embodiment the pharmaceutical composition comprises a compound offormula (I) together with a pharmaceutically acceptable carrier andoptionally one or more therapeutic agent(s).

In another embodiment the invention relates to the use of a combinationaccording to the invention in the manufacture of a pharmaceuticalcomposition for inhibiting the growth of tumour cells.

In a further embodiment the invention relates to a product containing acompound of formula (I) and one or more anticancer agent, as a combinedpreparation for simultaneous, separate or sequential use in thetreatment of patients suffering from cancer.

EXAMPLES

The invention will now be illustrated, but not limited, by reference tothe specific embodiments described in the following examples. Compoundsare named using an automated naming package such as AutoNom (MDL) or areas named by the chemical supplier.

The following synthetic procedures are provided for illustration of themethods used; for a given preparation or step the precursor used may notnecessarily derive from the individual batch synthesised according tothe step in the description given. In the examples, the followingabbreviations are used.

-   -   Ac₂O acetic anhydride    -   AcOH acetic acid    -   Boc tert-butyloxycarbonyl    -   Boc-Abu-OH (S)-2-(Boc-amino)butyric acid    -   BuLi butyllithium    -   CDI 1,1-carbonyldiimidazole    -   mCPBA m-chloroperbenzoic acid    -   DCM dichloromethane    -   DIPEA N-ethyl-N-(1-methylethyl)-2-propylamine    -   DMC dimethyl carbonate    -   DMF N,N-dimethylformamide    -   DMSO dimethyl sulfoxide    -   EDC 1-ethyl-3-(3′-dimethylaminopropyl)-carbodiimide        hydrochloride    -   Et₃N triethylamine    -   EtOAc ethyl acetate    -   EtOH ethanol    -   Et₂O diethyl ether    -   HATU 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium        hexafluorophosphate)    -   HBTU        O-benzotriazole-N,N,N′,N′-tetramethyl-uronium-hexafluoro-phosphate    -   HCl hydrochloric acid    -   HOAt 1-hydroxyazabenzotriazole    -   HOBt 1-hydroxybenzotriazole    -   HPLC high pressure liquid chromatography    -   IPA isopropyl alcohol    -   KHM DS potassium hexamethyldisilazide    -   LiHM DS lithium bis(trimethylsilyl)amide    -   MeCN acetonitrile    -   MeOH methanol    -   mins. minutes    -   MS mass spectrometry    -   NaBH(OAc)₃ sodium triacetoxyborohydride    -   NaOtBu potassium tert-butoxide    -   NMP N-methyl-2-pyrrolidinone    -   NMR nuclear magnetic resonance spectroscopy    -   oasfb on an anhydrous solvent free basis    -   Pd₂(dba)₃ tris(dibenzylideneacetone)dipalladium (0)    -   Pd(OAc)₂ palladium (2) acetate    -   Pd(PPh₃)₄ tetrakis(triphenylphosphine)palladium (0)    -   petrol petroleum ether fraction with boiling point range 40-60°        C.    -   PyBrop bromo-tris-pyrrolidino-phosphonium hexafluorophosphate    -   RT room temperature    -   SiO₂ silica    -   TBABr tetrabutylammonium bromide    -   TBAF tetrabutylammonium fluoride    -   TBME t-butylmethyl ether    -   TBTU N,N,N′,N′-tetramethyl-O-(benzotriazol-1-yl)uronium        tetrafluoroborate    -   TCNB 2,3,5,6-tetrachloronitrobenzene    -   TEA triethylamine    -   TFA trifluoroacetic acid    -   THF tetrahydrofuran    -   TMEDA N,N,N,N-tetramethylethylenediamine

NMR Data: Unless indicated, ¹H NMR spectra were recorded at 25° C. on aBruker Avance I spectrometer operating at 400 MHz. The data wereprocessed and analysed using Topspin 2.1 software. For NMR data, wherethe number of protons assigned is less than the theoretical number ofprotons in the molecule, it is assumed that the apparently missingsignal(s) is/are obscured by solvent and/or water peaks. In addition,where spectra were obtained in protic NMR solvents, exchange of NHand/or OH protons with solvent occurs and hence such signals arenormally not observed.

Analytical and Preparative LC-MS Systems

Analytical LC-MS system and method description

In the following examples, compounds were characterised by massspectroscopy using the systems and operating conditions set out below.Where atoms with different isotopes are present and a single massquoted, the mass quoted for the compound is the monoisotopic mass (i.e.³⁵Cl; ⁷⁹Br etc.).

Waters Platform LC-MS System:

-   HPLC System: Waters 2795-   Mass Spec Detector: Micromass Platform LC-   PDA Detector: Waters 2996 PDA    -   Platform MS conditions:-   Capillary voltage: 3.6 kV (3.40 kV on ES negative)-   Cone voltage: 30 V-   Source Temperature: 120° C.-   Scan Range: 125-800 amu-   Ionisation Mode: ElectroSpray Positive or    -   ElectroSpray Negative or    -   ElectroSpray Positive & Negative

Waters Fractionlynx LC-MS System:

-   HPLC System: 2767 autosampler-2525 binary gradient pump-   Mass Spec Detector: Waters ZQ-   PDA Detector: Waters 2996 PDA    -   Fractionlynx MS conditions:-   Capillary voltage: 3.5 kV (3.25 kV on ES negative)-   Cone voltage: 40 V (25 V on ES negative)-   Source Temperature: 120° C.-   Scan Range: 125-800 amu-   Ionisation Mode: ElectroSpray Positive or    -   ElectroSpray Negative or    -   ElectroSpray Positive & Negative

Agilent 1200SL-6140 LC-MS System—RAPID:

-   HPLC System: Agilent 1200 series SL-   Mass Spec Detector: Agilent 6140 single quadrupole-   Second Detector: Agilent 1200 MWD SL    -   Agilent MS conditions:-   Capillary voltage: 4000V on ES pos (3500V on ES Neg)-   Fragmentor/Gain: 100-   Gain: 1-   Drying gas flow: 7.0 L/min-   Gas Temperature: 345° C.-   Nebuliser Pressure: 35 psig-   Scan Range: 125-800 amu-   Ionisation Mode: ElectroSpray Positive-Negative switching

Preparative LC-MS System and Method Description

Preparative LC-MS is a standard and effective method used for thepurification of small organic molecules such as the compounds describedherein. The methods for the liquid chromatography (LC) and massspectrometry (MS) can be varied to provide better separation of thecrude materials and improved detection of the samples by MS.Optimisation of the preparative gradient LC method will involve varyingcolumns, volatile eluents and modifiers, and gradients. Methods are wellknown in the art for optimising preparative LC-MS methods and then usingthem to purify compounds. Such methods are described in Rosentreter U,Huber U.; Optimal fraction collecting in preparative LC/MS; J CombChem.; 2004; 6(2), 159-64 and Leister W, Strauss K, Wisnoski D, Zhao Z,Lindsley C., Development of a custom high-throughput preparative liquidchromatography/mass spectrometer platform for the preparativepurification and analytical analysis of compound libraries; J CombChem.; 2003; 5(3); 322-9.

Several systems for purifying compounds via preparative LC-MS aredescribed below although a person skilled in the art will appreciatethat alternative systems and methods to those described could be used.From the information provided herein, or employing alternativechromatographic systems, a person skilled in the art could purify thecompounds described herein by preparative LC-MS.

Waters Fractionlynx System:

-   -   Hardware:    -   2767 Dual Loop Autosampler/Fraction Collector    -   2525 preparative pump    -   CFO (column fluidic organiser) for column selection    -   RMA (Waters reagent manager) as make up pump    -   Waters ZQ Mass Spectrometer    -   Waters 2996 Photo Diode Array detector    -   Waters ZQ Mass Spectrometer    -   Waters MS running conditions:    -   Capillary voltage: 3.5 kV (3.2 kV on ES Negative)    -   Cone voltage: 25 V    -   Source Temperature: 120° C.    -   Scan Range: 125-800 amu    -   Ionisation Mode: ElectroSpray Positive or        -   ElectroSpray Negative

Agilent 1100 LC-MS Preparative System:

-   -   Hardware:    -   Autosampler: 1100 series “prepALS”    -   Pump: 1100 series “PrepPump” for preparative flow gradient and        1100 series “QuatPump”    -   for pumping modifier in prep flow    -   UV detector: 1100 series “MWD” Multi Wavelength Detector    -   MS detector: 1100 series “LC-MSD VL”    -   Fraction Collector: 2×“Prep-FC”    -   Make Up pump: “Waters RMA”    -   Agilent Active Splitter    -   Agilent MS running conditions:    -   Capillary voltage: 4000 V (3500 V on ES Negative)    -   Fragmentor/Gain: 150/1    -   Drying gas flow: 12.0 L/min    -   Gas Temperature: 350° C.    -   Nebuliser Pressure: 50 psig    -   Scan Range: 125-800 amu    -   Ionisation Mode: ElectroSpray Positive or        -   ElectroSpray Negative    -   Columns:

A range of commercially available columns—both achiral and chiral—may beused such that, in conjunction with the changes in mobile phase, organicmodifier and pH, they enabled the greatest cover in terms of a broadrange of selectivity. All columns were used in accordance with themanufacturers recommended operating conditions. Typically 5 micronparticle sized columns were used where available. For example, columnsfrom Waters (including but not limited to XBridge™ Prep OBD™ C18 andPhenyl, Atlantis® Prep T3 OBD™ and Sunfire™ Prep OBD C18 5 μm 19×100mm), Phenomenex (including but not limited to Synergy MAX-RP and LUX™Cellulose-2), Astec (Chirobiotic™ columns including but not limited toV, V2 and T2) and Diacel® (including but not limited to Chiralpak® AD-H)were available for screening.

-   -   Eluents:

Mobile phase eluent was chosen in conjunction with column manufacturersrecommended stationary phase limitations in order to optimise a columnsseparation performance.

-   -   Methods:

Achiral Preparative Chromatography

The compound examples described have undergone HPLC purification, whereindicated, using methods developed following recommendations asdescribed in Snyder L. R., Dolan J. W., High-Performance GradientElution The Practical Application of the Linear-Solvent-Strength Model,Wley, Hoboken, 2007.

Chiral Preparative Chromatography

Preparative separations using Chiral Stationary Phases (CSPs) are thenatural technique to apply to the resolution of enantiomeric mixtures.Equally, it can be applied to the separation of diastereomers andachiral molecules. Methods are well known in the art for optimisingpreparative chiral separations on CSPs and then using them to purifycompounds. Such methods are described in Beesley T. E., Scott R. P. W.;Chiral Chromatography; Wiley, Chichester, 1998.

The values of salt stoichiometry or acid content in the compounds asprovided herein, are those obtained experimentally and may varydependent on the analytical method used. In case no salt form isindicated, the compound was obtained as a free base.

Preparation 1:(R)-2-((S)-2-Benzyloxycarbonylamino-3-hydroxy-propionyl-amino)-propionicacid methyl ester

Diisopropylethylamine (375 mL) was added dropwise to a cooled mixture of(R)-2-amino-propionic acid methyl ester hydrochloride (100 g, 0.716mol), EDC (165 g, 0.86 mol), carbobenzyloxy-L-serine (171.4 g, 0.716mol) and DCM (3.6 L). The resulting mixture was stirred under nitrogenat ambient temperature for 16 h. After removing solvent in vacuo at 40°C., the residue was diluted with saturated sodium carbonate (1 L), water(1 L) and extracted with EtOAc (2 L, 2×1 L). The combined organic phaseswere washed with 2 M hydrochloric acid (1 L), saturated brine solution(1 L), dried over magnesium sulfate and concentrated in vacuo at 40° C.,to give the title compound (172 g) as a colourless solid. ¹H NMR(Me-d3-OD): 7.44-7.28 (6H, m), 5.13 (2H, s), 4.46 (1H, d), 4.43 (1H, d),4.25 (1H, t), 3.82-3.68 (5H, m), 1.39 (3H, d).

Preparation 2: (3S,6R)-3-Hydroxymethyl-6-methyl-piperazine-2,5-dione

To(R)-2-((S)-2-benzyloxycarbonylamino-3-hydroxy-propionyl-amino)-propionicacid methyl ester (which may be prepared as described in Preparation 1)(172 g, 0.53 mol) was added 10% palladium on carbon (8.6 g), MeOH (530mL) and cyclohexene (344 mL) under nitrogen. The mixture was heated toreflux for 17 h. MeOH (500 mL) was added and the reflux continued for 1h. The hot reaction mixture was filtered through a pad of celite, cakewashing with hot MeOH (2×500 mL). The combined filtrates wereconcentrated. The resulting solid was slurried in 2-butanone (400 mL)and petrol (400 mL) was added gradually over 10 min. After stirring for30 min, the solids were filtered, cake washed with 2:1 petrol/2-butanone(300 mL). The filter cake was dried in vacuo at 40° C., to give thetitle compound (68.3 g) as an off white solid. ¹H NMR (DMSO-d6): 8.08(1H, s), 7.90 (1H, s), 5.11 (1H, t), 3.92 (1H, q), 3.80-3.71 (1H, m),3.71-3.60 (1H, m), 3.58-3.47 (1H, m), 1.24 (3H, d).

Preparation 3: ((2R,5R)-5-Methyl-piperazin-2-yl)-methanol hydrochloride

To (3S,6R)-3-hydroxymethyl-6-methyl-piperazine-2,5-dione (which may beprepared as described in Preparation 2) (34 g, 0.215 mol) was added asolution of borane in THF (1 M, 1.6 L, 1.6 mol) and the mixture washeated to 70° C. for 18 h. The solution was cooled in ice, then MeOH(425 mL) was gradually added, followed by 5 M hydrochloric acid (113mL). The mixture was heated to 70° C. for 2 h and then cooled to ambienttemperature. The resulting solid was filtered, cake washed with THF (200mL) and dried in vacuo at 40° C., to give the title compound (39.3 g) asa colourless solid. ¹H NMR (DMSO-d6): 9.79 (3H, s), 5.59 (1H, s),3.76-3.40 (5H, m), 3.19-2.94 (2H, m), 1.28 (3H, d).

Preparation 4: (2R,5R)-5-Hydroxymethyl-2-methyl-piperazine-1-carboxylicacid tert-butyl ester

To ((2R,5R)-5-methyl-piperazin-2-yl)-methanol hydrochloride (which maybe prepared as described in Preparation 3) (20 g, 119 mmol) in MeOH (96mL) at 0° C. (ice bath) was added triethylamine (48.7 mL, 357 mmol).tert-Butyl dicarbonate (61 g, 280 mmol) in MeOH (145 mL) was added over30 min. The reaction temperature was maintained at <10° C. for 1 h,warmed to ambient temperature over 1 h and then heated to 50° C. for 18h. The reaction was concentrated and the residue dissolved in ethanol(397 mL). A solution of NaOH (23.8 g, 595 mmol) in water (397 mL) wasadded and the reaction heated to 100° C. for 18 h, then cooled toambient temperature. Mixture was neutralised with 1 M HCl (˜300 mL) topH 9 (using a pH meter), then extracted with chloroform (3×700 mL),dried over sodium sulfate, filtered and concentrated. The residue wasredissolved in MeOH and concentrated, then dried in vacuo at 40° C., togive the title compound (21 g, 75%) as a colourless solid. ¹H NMR(Me-d3-OD): 4.20-4.07 (1H, m), 3.79 (1H, dd), 3.71-3.58 (2H, m), 3.54(1H, dd), 3.24 (1H, dd), 3.18-3.01 (1H, m), 3.01-2.89 (1H, m), 2.55 (1H,dd), 1.48 (9H, s), 1.25 (3H, s).

Preparation 5:(2R,5R)-4-Benzyl-5-hydroxymethyl-2-methyl-piperazine-1-carboxylic acidtert-butyl ester

A mixture of (2R,5R)-5-hydroxymethyl-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation4) (3.48 g, 15.1 mmol), benzaldehyde (1.76 g, 16.6 mmol), sodiumtriacetoxyborohydride (3.84 g, 18.1 mmol) and 1,2-dichloroethane (30 mL)was stirred at 20° C. for 18 h, then partitioned between saturatedaqueous NaHCO₃ (150 mL) and DCM (3×50 mL). Combined organic extractswere dried (Na₂SO₄) then evaporated in vacuo to give an oil.Chromatography (SiO2, 0-30% EtOAc in petrol) gave the title compound(4.588 g, 74%) as a colourless solid. MS: [M+H]⁺=321.

Preparation 6:(2R,5R)-4-Benzyl-5-chloromethyl-2-methyl-piperazine-1-carboxylic acidtert-butyl ester

Methanesulfonyl chloride (570 μL, 7.35 mmol) was added to a solution of(2R,5R)-4-benzyl-5-hydroxymethyl-2-methyl-piperazine-1-carboxylic acidtert-butyl ester (which may be prepared as described in Preparation 5)(1.9 g, 6.12 mmol) containing TEA (2.6 mL, 18.4 mmol) in DCM (30 mL) at0° C. The solution was stirred at room temperature for 18 h. Thereaction was partitioned between aqueous NH₄Cl and DCM. The organicphase was collected, dried over MgSO4, filtered and concentrated invacuo. Chromatography (30% EtOAc in petrol) gave the title compound (1.6g) as a white solid. MS: [M+H]⁺=339.

Preparation 7:(2R,5S)-4-Benzyl-2-methyl-5-((R)-3-methyl-morpholin-4-ylmethyl)-piperazine-1-carboxylicacid tert-butyl ester

K₂CO₃ (81.6 g, 591 mmol) and KI (73.6 g, 443 mmol) were added to asolution of(2R,5R)-4-benzyl-5-chloromethyl-2-methyl-piperazine-1-carboxylic acidtert-butyl ester (which may be prepared as described in Preparation 6)(50 g, 147.9 mmol) in acetonitrile (400 mL) followed by(R)-3-methyl-morpholine hydrochloride (26.4 g, 192 mmol). The reactionwas stirred at 70° C. for 18 h. The solid was then removed by filtrationand the solvent removed in vacuo. The crude material was purified bychromatography using a pad of silica (20% EtOAc in Petrol) to give thetitle compound (41.3 g) as a white solid. MS: [M+H]⁺=404.

Preparation 8:(2R,5S)-2-Methyl-5-((R)-3-methyl-morpholin-4-ylmethyl)-piperazine-1-carboxylicacid tert-butyl ester

Palladium on carbon (10%) (33 g) and acetic acid (220 mL) were added toa solution of(2R,5S)-4-benzyl-2-methyl-5-((R)-3-methyl-morpholin-4-ylmethyl)-piperazine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation7) (41.3 g, 102 mmol) in EtOH (300 mL). The mixture was stirred under H₂(1 atmosphere) at room temperature for 18 h. The reaction mixture wasthen filtered through a pad of Celite to remove the catalyst and thesolvent was removed in vacuo. The crude material was partitioned betweensaturated aqueous NaHCO₃ and DCM and the product extracted with DCM(3×). The organic phase was dried over MgSO₄, filtered and concentratedin vacuo to give the title compound (30.5 g) as a pale yellow oil. ¹HNMR (400 MHz, CDCl3): 4.43-3.87 (1H, m), 3.78 (1H, d), 3.73-3.55 (3H,m), 3.32 (1H, dd), 3.22 (1H, dd), 3.16-2.93 (3H, m), 2.93-2.72 (1H, m),2.55-2.35 (2H, m), 2.35-2.15 (2H, m), 1.89 (1H, dd), 1.45 (9H, s), 1.26(3H, d), 0.96 (3H, d).

Alternative Procedure:

To a tightly sealed 10 L flange flask fitted with a stirrer bar wasadded(2R,5S)-4-benzyl-2-methyl-5-((R)-3-methyl-morpholin-4-ylmethyl)-piperazine-1-carboxylicacid tert-butyl ester (500 g, 1.24 mol, 1.0 eq) (which may be preparedas described in Preparation 7) and ethanol (Stock, 5 L). The flask wasplaced under nitrogen and 10% Pd/C (Aldrich, 50 g, 0.124 mol, 0.1 eq)was added as a paste in ethanol. The flask was purged several times witha di-vac pump and placed under a hydrogen atmosphere using 4 balloons.The reaction was warmed to 30° C. overnight after which time NMRconfirmed complete consumption of starting material. The reactionmixture was cooled to room temperature and filtered through a pad ofcelite under nitrogen. The filtrates were evaporated to dryness toafford the title product as a colourless oil.

¹H NMR (MeOD): 1.00 (3H, d), 1.25 (3H, d), 1.48 (9H, s), 2.08-2.14 (1H,m), 2.28-2.35 (1H, m), 2.42-2.48 (1H, m), 2.49-2.55 (1H, dd), 2.80-3.06(4H, m), 3.22-3.28 (2H, m), 3.61-3.78 (4H, m), 4.12-4.16 (1H, m).

¹³C NMR (MeOD): 14.6, 15.7, 28.8, 40.8, 44.8, 48.3, 50.3, 53.2, 54.3,57.5, 68.5, 73.9, 81.1, 157.0.

Preparation 9:(2R,5S)-4-Benzyl-5-((3R,5R)-3,5-dimethyl-morpholin-4-ylmethyl)-2-methyl-piperazine-1-carboxylicacid tert-butyl ester

K₂CO₃ (2.7 g, 19.5 mmol) and KI (1.83 g, 11.05 mmol) were added to asolution of(2R,5R)-4-benzyl-5-chloromethyl-2-methyl-piperazine-1-carboxylic acidtert-butyl ester (which may be prepared as described in Preparation 6)(2.2 g, 6.5 mmol) in acetonitrile (30 mL) followed by (3R,5R)-3,5-dimethyl-morpholine (0.80 g, 7.0 mmol). The reaction was stirredat 70° C. for 18 h. The solid was then removed by filtration and thesolvent removed in vacuo. The residue was partitioned between water anddichloromethane. The organic phase was dried, filtered and the solventevaporated. The crude material was purified by chromatography on silica(0-40% EtOAc in Petrol) to give the title compound (2.56 g, 94%) as awhite solid. MS: [M+H]⁺=418.

Preparation 10:(2R,5S)-5-((3R,5R)-3,5-Dimethyl-morpholin-4-ylmethyl)-2-methyl-piperazine-1-carboxylicacid tert-butyl ester

Palladium on carbon (10%) (1.6 g) and acetic acid (10 mL) were added toa solution of(2R,5S)-4-benzyl-5-((3R,5R)-3,5-dimethyl-morpholin-4-ylmethyl)-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation9) (2.5 g, 6.0 mmol) in EtOH (70 mL). The mixture was stirred under H2(1 atmosphere) at room temperature for 3 h. The reaction mixture wasthen filtered through a pad of Celite to remove the catalyst and thesolvent was removed in vacuo. The crude material was partitioned betweensaturated aqueous NaHCO₃ and DCM and the product extracted with DCM(3×). The organic phase was dried over MgSO₄, filtered and concentratedin vacuo to give the title compound (1.53 g, 78%) as a pale yellow oil.¹H NMR (400 MHz, CDCl3): 4.16 (1H, s), 3.79-3.59 (3H, m), 3.44-3.19 (3H,m), 3.08 (1H, dd), 2.99-2.69 (4H, m), 2.52 (1H, dd), 2.29 (1H, dd), 1.47(9H, s), 1.27 (3H, d), 1.00 (6H, d).

The following compound was made following an analogous procedure to thatdescribed in Preparations 9 and 10:

-   10A:    (2R,5S)-5-((2S,5R)-2,5-Dimethyl-morpholin-4-ylmethyl)-2-methyl-piperazine-1-carboxylic    acid tert-butyl ester, MS: [M+H]⁺=328.

Preparation 11:2-(5-Chloro-3-fluoro-pyridin-2-yl)-2-methyl-propionitrile

A solution of sodium bis(trimethylsilyl)amide (610 mL, 40% intetrahydrofuran, 1.326 mole) was added to an ice-cooled solution of5-chloro-2,3-difluoropyridine (198.2 g, 1.326 mole) and isobutyronitrile(238 mL, 2.65 mole) in toluene (2 L). The mixture was stirred undernitrogen at RT overnight before addition of saturated aqueous ammoniumchloride (1 L). Phases were separated and the aqueous extracted withethyl acetate (2×1 L). Combined organic extracts were dried (MgSO₄) andconcentrated in vacuo at 40° C. to give the title compound (259.8 g,95%)¹H NMR (400 MHz, DMSO-d6): 8.57 (1H, dd), 8.24 (1H, dd), 1.74 (6H,broad).

Preparation 12: 2-(5-Chloro-3-fluoropyridin-2-yl)-2-methylpropylamine

Borane-tetrahydrofuran complex (1 M, 1.37 L, 1.365 mole) was added to acooled solution of2-(5-chloro-3-fluoro-pyridin-2-yl)-2-methyl-propionitrile (which may beprepared as described in Preparation 11) (135.6 g, 0.683 mole) intetrahydrofuran (670 mL). The mixture was stirred under nitrogen at roomtemperature overnight before cooling in ice. The mixture was quenched bythe addition of 5 M hydrochloric acid (335 mL). The resulting mixturewas basified with 40% aqueous potassium hydroxide (460 mL) and thephases were separated. The basic aqueous phase was extracted with ethylacetate (2×670 mL) and the combined organic extracts were washed withbrine (670 mL), dried (MgSO₄) and concentrated in vacuo at 40° C. togive the title compound (102.9 g, 74%)¹H NMR (400 MHz, DMSO-d6): 8.44(1H, t), 7.95 (1H, dd), 2.85 (2H, d), 1.29 (6H, d).

Preparation 12, alternative procedure:2-(5-Chloro-3-fluoropyridin-2-yl)-2-methylpropylamine

To a 10 L flange flask was added2-(5-chloro-3-fluoro-pyridin-2-yl)-2-methyl-propionitrile (which may beprepared as described in Preparation 11) (200 g, 1.00 mol), nickel(II)chloride hexahydrate (239.4 g, 1.00 mol) and ethanol (3.0 L). Theresulting green solution was cooled to 0° C. using a dry ice/acetonebath under an atmosphere of nitrogen. Sodium borohydride (114.3 g, 3.02mol) was added portionwise at such a rate that the reaction temperatureremained below 6° C. (addition time=1% h) to give a black suspension.Once the addition was complete, the cold bath was replaced with anice/water bath, then the reaction was allowed to warm to RT overnight.The reaction mixture was cooled to 0-4° C. in an ice bath. 25% Aqueousammonia solution (2680 mL) was added from a dropping funnel such thatthe reaction temperature remained below 10° C. (addition time=1 h). Onceaddition was complete, stirring was continued at ca 0° C. for 30 minthen the mixture was filtered through celite, and the residues washedwith ethanol (2×750 mL). (Care! Don't let the filter pad dry up. Totalfiltration time ca 2 h.) The pale yellow/brown filtrate was transferredto a large rotary evaporator and concentrated until all ethanol had beenremoved. The resulting green oil was transferred to a 5 L separatingfunnel and 25% aqueous ammonia solution added until the oil turnedyellow (200 mL). The oil was separated and the aqueous phase extractedwith toluene (2×300 mL). The combined organic extracts were washed with1:1 25% aqueous ammonia solution/brine (300 mL), dried over sodiumsulfate, filtered and concentrated on the rotary evaporator (bathtemperature up to 70° C.) to give the crude product as a yellow oil (161g), data consistent with those given above. This was used in the nextstep without purification.

Preparation 13:6-Chloro-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine

A mixture of 2-(5-chloro-3-fluoropyridin-2-yl)-2-methylpropylamine(which may be prepared as described in Preparation 12 and Preparation12, alternative procedure) (33 g, 0.163 mole), potassium carbonate (122g, 0.884 mole) and NMP (100 mL) was heated to 150° C. for 4 hours. Thecooled mixture was diluted with water (330 mL) and extracted withtoluene (3×300 mL) The combined organic extracts were washed with brine(160 mL), dried (MgSO₄) and concentrated in vacuo at 40° C. to givecrude material (24.8 g). Chromatography on silica eluting with 5-30%ethyl acetate/petrol gave the title compound (21 g, 71%)¹H NMR (400 MHz,DMSO-d6): 7.61 (1H, d), 6.75 (1H, d), 6.06 (1H, bs), 3.31 (2H, s), 1.21(6H, s).

Preparation 14:6-Chloro-3,3-dimethyl-2,3-dihydropyrrolo[3,2-b]pyridine-1-carboxylicacid tert-butyl ester

Di-tertbutyldicarbonate (3.7 g, 17.1 mmol) was added to a mixture of6-chloro-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine (which maybe prepared as described in Preparation 13) (2.6 g, 14.2 mmol),tetrahydrofuran (26 mL) and 2 M sodium hydroxide (11.4 mL, 22.8 mmol)with stirring over 2 days. The biphasic mixture was diluted with water(20 mL) and extracted with ethyl acetate (2×20 mL). The combined organicextracts were dried (MgSO₄) and concentrated in vacuo at 40° C. to givecrude material (6.02 g). Chromatography on silica eluting with 5-30%ethyl acetate/petrol gave the title compound (2.23 g, 55%); ¹H NMR (400MHz, DMSO-d6): 8.11 (1H, d), 7.85 (1H, bs), 3.77 (2H, s), 1.52 (9H, s),1.28 (6H, s).

Preparation 15:6-(4-Fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine

A solution of 4-fluorobenzylzinc chloride (2 L of 0.5 M solution in THF,1 mol) was added to a degassed mixture of6-chloro-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine (which maybe prepared as described in Preparation 13) (91.3 g, 0.5 mol), lithiumbromide (130.3 g, 1.5 mol) and[1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(11)dichloride (6.8 g, 0.01 mol) in THF (685 mL) and NMP (910 mL) at 20° C.with exotherm. The resulting dark mixture was stirred under nitrogen atroom temperature for 18 h. The reaction was quenched with 2.5% aqueouscitric acid (900 mL) and extracted with toluene (2×900 mL). The combinedorganic phases were washed with water (3×900 mL), brine (900 mL), driedover MgSO4, filtered and concentrated in vacuo. The resulting solid wasslurried in petrol (450 mL) and toluene (100 mL). After stirring for 30min, the solids were filtered, cake washed with petrol (2×90 mL). Thefilter cake was dried in vacuo at 40° C., to give the title compound(107.3 g) as a grey solid. ¹H NMR (DMSO-d6): 7.60 (1H, d), 7.30-7.22(2H, m), 7.15-7.06 (2H, m), 6.53 (1H, d), 5.64 (1H, s), 3.78 (2H, s),3.22 (2H, d), 1.19 (6H, s).

The following compounds were prepared in a similar manner to thatdescribed in Preparation 15:

15A: tert-Butyl6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridine-1-carboxylate,MS: [M+H]⁺=357.

15B:6-(3-Fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine,MS: [M+H]⁺=257.

15C: 6-Butyl-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine, MS:[M+H]⁺=205.

15D:6-(2-Fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine,MS: [M+H]⁺=257.

15E:6-(2,4-Difluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine

Preparation 16:5-Bromo-6-(4-fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine

A solution of6-(4-fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine(which may be prepared as described in Preparation 15) (88.5 g, 0.345mol) in DMF (1.67 L) was cooled to −5° C. Solid N-bromosuccinimide (61.5g, 0.345 mol) was added in portions with exotherm. The mixture wasstirred for 1 h warming to room temperature. Water (2.66 L) was addedwith exotherm and the resulting mixture was stirred for 18 h at roomtemperature The solids were filtered and cake washed with water (270mL). The filter cake was dissolved in THF (1.5 L), dried over MgSO4,filtered and concentrated in vacuo to give the title compound (109.7 g)as a yellow solid. ¹H NMR (DMSO-d6): 7.29-7.20 (2H, m), 7.20-7.03 (2H,m), 6.64 (1H, s), 5.88 (1H, s), 3.89 (2H, s), 3.26 (2H, d), 1.20 (6H,s).

The following compounds were prepared in a similar manner to Preparation16:

16A:5-Bromo-6-(3-fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine,MS: [M+H]⁺=335, 337.

16B:5-Bromo-6-(2-fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine,MS: [M+H]⁺=335, 337.

16C: 5-Bromo-6-Butyl-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine,MS: [M+H]⁺=283, 285.

16D:5-Bromo-6-(2,4-difluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine

Preparation 17:[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-methanol

To5-bromo-6-(4-fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine(which may be prepared as described in Preparation 16) (22.8 g, 68.2mmol) in THF (300 mL), cooled to −78° C., was added MeLi (1.6 M in Et₂O;51.1 mL, 91.8 mmol) over 15 minutes. tert-Butyllithium (1.7 M in hexane;96 mL,164 mmol) was then added over 30 minutes. After 15 minutes, DMF(26 mL) was added and the mixture stirred at −78° C. for a further 50minutes. Saturated aqueous NH₄Cl (450 mL) was added and the mixture wasstirred for 10 minutes at RT. The organic layer was isolated and theaqueous layer extracted with EtOAc (2×150 mL). The combined organicfractions were washed with brine (200 mL), dried (MgSO4) and evaporatedto give6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-5-carbaldehydeas a yellow solid which was used without any further purification. MS:m/z=285 (M+H+)+. This product (˜68 mmol) was suspended in MeOH (250 mL)and cooled in an ice bath. NaBH₄ (3.4 g, 81.8 mmol) was addedportion-wise over 5 minutes. Cooling was removed and the mixture stirredfor a further 20 minutes. The mixture was cooled in an ice bath followedby careful addition of 10% aqueous KHSO₄ over 10 minutes (care:effervescence). After stirring for 5 minutes at RT the mixture wasre-cooled using an ice bath. The mixture was basified by addition of 50%aqueous NaOH (˜18 mL) and then concentrated in vacuo to—one thirdvolume. The resulting aqueous mixture was extracted with CH₂Cl₂ (1×200mL, 2×100 mL) and the combined CH₂Cl₂ layers were dried (MgSO₄). TheCH₂Cl₂ solution was concentrated in vacuo to ˜30 mL and then dilutedwith toluene (70 mL) to initiate crystallisation of the product.Collection by filtration gave the product as a colourless crystallinesolid (10.6 g). A second crop (2.1 g) was collected from the filtrate.The filtrate was concentrated and the remaining material was purified bySiO2 chromatography (eluting with 25-50% EtOAc/hexanes) to give a thirdbatch of material (2.1 g); giving the title compound in an overall yieldof 14.8 g (76% over 2 steps), MS: [M+H]⁺=287. An alternative procedureinvolves subsequent recrystallization from isopropyl alcohol.

The following compound was prepared in a similar manner to Preparation17:(6-Butyl-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl)-methanol,MS: [M+H]⁺=235.

Preparation 18:2-Chloro-1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}ethan-1-one

To a cooled (˜5° C.) suspension of[6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-methanol(which may be prepared as described in Preparation 17) (11.8 g, 41.3mmol) in MeCN (175 mL) was added chloroacetyl chloride (6.9 mL, 86.7mmol). Cooling was removed and the mixture stirred for 30 minutes at RT.The mixture was then evaporated in vacuo and dissolved in MeOH (200 mL).K₂CO₃ solution (12 g in 100 mL H₂O) was added and the mixture stirred atRT for 20 minutes after which the mixture was concentrated in vacuo toone quarter volume. The aqueous mixture was extracted with CH₂Cl₂ (1×100mL, 2×30 mL) and the combined CH₂Cl₂ layers were dried (MgSO₄).Evaporation in vacuo gave the product as colourless crystalline solid(12.1 g,˜100%), MS: [M+H]⁺=363.

The following compounds were prepared following a method analogous orsimilar to that of Preparation 18:

18A:1-(2-Chloroacetyl)-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=349.

18B:2-Chloro-1-{6-[(2-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}ethan-1-one,MS: [M+H]⁺=363.

18C:1-(2-Chloroacetyl)-6-[(4-fluorophenyl)methyl]-3,3,4-trimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=363.

18D:1-(2-Chloroacetyl)-6-[(2,4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=367.

18E:1-(2-Chloroacetyl)-6-[(2-fluorophenyl)methyl]-3,3,4-trimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=363.

18F:2-Chloro-1-{6-[(2,4-difluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}ethan-1-one,MS: [M+H]⁺=381.

18G:2-Chloro-1-[5-(1,2-dihydroxyethyl)-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl]ethan-1-one,MS: [M+H]⁺=393.

18H:2-Chloro-1-{6-[(3-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}ethan-1-one,MS: [M+H]⁺=363.

18I:1-(2-Chloro-acetyl)-6-(4-fluoro-benzyl)-3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo[2,3-c]pyridin-5-one,MS: [M+H]⁺=349.

18J:1-(2-Chloro-acetyl)-6-(2,4-difluoro-benzyl)-3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo[2,3-c]pyridin-5-one,MS: [M+H]⁺=367.

18K: 2-Chloro-1-[5-((R orS)-1,2-dihydroxyethyl)-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl]ethan-1-one,from slower eluting precursor. MS: [M+H]⁺=393.

18L:1-(2-Chloroacetyl)-6-[(2,4-fluorophenyl)methyl]-3,3,4-trimethyl-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=381.

18M: 2-Chloro-1-{6-[(4-fluorophenyl)methyl]-5-(R orS)-1-hydroxy-2-methoxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}ethan-1-one,from faster eluting precursor, MS: [M+H]⁺=407.

18N: 2-Chloro-1-{6-[(4-fluorophenyl)methyl]-5-(R orS)-1-methoxy-2-hydroxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}ethan-1-one,from faster eluting precursor, MS: [M+H]⁺=407.

18O: 2-Chloro-1-{6-[(4-fluorophenyl)methyl]-5-(R orS)-1-hydroxy-2-methoxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}ethan-1-one,from slower eluting precursor, MS: [M+H]⁺=407.

18P: 2-Chloro-1-{6-[(4-fluorophenyl)methyl]-5-(R orS)-1-methoxy-2-hydroxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}ethan-1-one,from slower eluting precursor, MS: [M+H]⁺=407.

18Q:1-(2-Chloro-acetyl)-6-(2,4-difluoro-benzyl)-3,3,4-trimethyl-1,2,3,6-tetrahydro-pyrrolo[2,3-c]pyridin-5-one,MS: [M+H]⁺=381.

18R:6-Butyl-1-(2-chloro-acetyl)-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=297.

18S:1-[6-Butyl-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl]-2-chloroethan-1-one,MS: [M+H]⁺=311.

18T:6-Butyl-1-(2-chloro-acetyl)-3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo[2,3-c]pyridin-5-one,MS: [M+H]⁺=297.

Preparation 19: tert-Butyl(2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate

(2R,5S)-2-Methyl-5-((R)-3-methyl-morpholin-4-ylmethyl)-piperazine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation8) (15.5 g, 46.4 mmol), KI (12.8 g, 77.4 mmol) and K₂CO₃ (21.4 g, 155mmol) were stirred in MeCN (70 mL) and cooled in an ice bath.2-Chloro-1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}ethan-1-one(which may be prepared as described in Preparation 18) (14.0 g, 38.7mmol) was then added as a solution in MeCN (100 mL). The mixture wasstirred at RT for 2 hours and then concentrated in vacuo to ˜one quartervolume. The mixture was partitioned between EtOAc (150 mL) and H₂O (150mL) and then the aqueous layer extracted with further EtOAc (1×75 mL).The combined EtOAc layers were washed with 10% aqueous KH₂PO₄ (4×100 mL)and then brine (70 mL). The organic layer was dried (MgSO₄) andevaporated to give the product as a colourless solid (25.8 g, 98%), MS:[M+H]⁺=640.

The following compounds were prepared following a method analogous tothat of Preparation 19:

tert-Butyl(2R,5S)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-4-(2-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=654.

tert-Butyl(2R,5S)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-4-(2-{6-[(4-fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=640.

tert-Butyl(2R,5S)-4-(2-{6-[(2-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=640.

tert-Butyl(2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=626.

tert-Butyl(2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-3,3,4-trimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=640.

tert-Butyl(2R,5S)-4-(2-{6-[(2,4-difluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=658.

tert-Butyl(2R,5S)-4-(2-{6-[(2,4-difluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=644.

tert-Butyl(2R,5S)-4-(2-{6-[(2-fluorophenyl)methyl]-3,3,4-trimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=640.

tert-Butyl(2R,5S)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-4-(2-{6-[(4-fluorophenyl)methyl]-3,3,4-trimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=654.

tert-Butyl(2R,5S)-4-(2-{6-[(2,4-difluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,1H NMR (400 MHz, Me-d3-OD): 8.12 (1H, s), 7.27-7.16 (1H, m), 7.06-6.86(2H, m), 4.76 (2H, s), 4.17 (1H, s), 4.10-4.07 (2H, m), 3.99 (1H, d),3.74-3.49 (5H, m), 3.30-3.22 (2H, m), 2.97-2.77 (4H, m), 2.59-2.43 (2H,m), 2.43-2.32 (1H, m), 2.32-2.21 (1H, m), 1.47 (9H, s), 1.43 (6H, s),1.22 (3H, d), 1.00 (3H, d).

tert-Butyl(2R,5S)-4-{2-[5-(1,2-dihydroxyethyl)-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl]-2-oxoethyl}-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylateMS: [M+H]⁺=670; chiral HPLC (heptane/ethanol, 80:20, 0.2% DEA,chiralPAk-IC column) gave faster eluting diastereoisomer A, MS:[M+H]⁺=670 and slower eluting diastereoisomer B, MS: [M+H]⁺=670.

tert-Butyl(2R,5S)-4-(2-{6-[(3-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=640.

tert-Butyl(2R,5S)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-4-(2-{6-[(4-fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-1-yl}-2-oxoethyl)-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=640.

tert-Butyl(2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=626.

tert-Butyl(2R,5S)-5-{[(2S,5R)-2,5-dimethylmorpholin-4-yl]methyl}-4-(2-{6-[(4-fluorophenyl)methyl]-3,3,4-trimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=654.

tert-Butyl(2R,5S)-4-(2-{6-[(2,4-difluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-1-yl}-2-oxoethyl)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=658.

tert-Butyl (2R,5S)-4-{2-[5-((R orS)-1,2-dihydroxyethyl)-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl]-2-oxoethyl}-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=684.

tert-Butyl(2R,5S)-4-(2-{6-[(2,4-difluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-1-yl}-2-oxoethyl)-5-{[(2S,5R)-2,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=658.

tert-Butyl(2R,5S)-4-(2-{4-amino-6-[(2,4-difluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=659.

tert-Butyl(2R,5S)-4-(2-{4-amino-6-[(4-fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=641.

tert-Butyl(2R,5S)-4-(2-{6-[(2,4-difluorophenyl)methyl]-3,3,4-trimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-5-{[(2S,5R)-2,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=672.

tert-Butyl (2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-5-((R orS)1-hydroxy-2-methoxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate(from faster eluting isomer), MS: [M+H]⁺=684.

tert-Butyl (2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-5-((R orS)₁-hydroxy-2-methoxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate(from slower eluting isomer), MS: [M+H]⁺=684.

tert-Butyl (2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-5-((R orS)₁-methoxy-2-hydroxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate(from faster eluting isomer), MS: [M+H]⁺=684.

tert-Butyl (2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-5-((R orS)₁-methoxy-2-hydroxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate(from slower eluting isomer), MS: [M+H]⁺=684.

tert-Butyl(2R,5S)-4-(2-{4-amino-6-butyl-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=603.

tert-Butyl(2R,5S)-4-(2-{6-[(2,4-difluorophenyl)methyl]-3,3,4-trimethyl-5-oxo-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate,MS: [M+H]⁺=658.

tert-Butyl(2R,5S)-4-(2-{6-butyl-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=588.

tert-Butyl(2R,5S)-4-(2-{6-butyl-3,3,4-trimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=602.

tert-Butyl(2R,5S)-4-{2-[6-butyl-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl]-2-oxoethyl}-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=602.

tert-Butyl(2R,5S)-4-(2-{6-butyl-3,3-dimethyl-5-oxo-1H,2H,3H,5H,6H-pyrrolo[2,3-c]pyridin-1-yl}-2-oxoethyl)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=588.

tert-Butyl(2R,5S)-5-{[(3R,5R)-3,5-dimethylmorpholin-4-yl]methyl}-4-(2-{6-[(4-fluorophenyl)methyl]-5-((RorS)-2-hydroxy-1-methoxyethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=698.

tert-Butyl(2R,5S)-4-(2-{6-butyl-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-5-{[(2S,5R)-2,5-dimethylmorpholin-4-yl]methyl}-2-methylpiperazine-1-carboxylate,MS: [M+H]⁺=588.

Preparation 20:(2R,5S)-4-{2-[6-(2,4-Difluoro-benzyl)-3,3,4-trimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo[3,2-b]pyridin-1-yl]-2-oxo-ethyl}-5-((3R,5R)-3,5-dimethyl-morpholin-4-ylmethyl)-2-methyl-piperazine-1-carboxylicacid tert-butyl ester

(2R,5S)-4-{2-[6-(2,4-Difluoro-benzyl)-3,3-dimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo[3,2-b]pyridin-1-yl]-2-oxo-ethyl}-5-((3R,5R)-3,5-dimethyl-morpholin-4-ylmethyl)-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (146 mg, 0.22 mmol) was dissolved in DMF (3 mL).Sodium hydride (60%, 11 mg, 0.27 mmol) was added and the reactionmixture was stirred for 30 mins. Iodomethane (0.017 mL, 0.27 mmol) wasadded and the reaction was stirred for 30 mins at room temperaturebefore being partitioned between water (10 mL) and EtOAc (2×10 mL). Theorganic fractions were washed with brine, dried over magnesium sulfateand concentrated. The residue was purified by column chromatography,eluting with 0-10% MeOH in EtOAc and then by preparative HPLC to givethe title compound (17.6 mg). MS: [M+H]⁺=672.

Preparation 21:(2R,5S)-4-{2-[6-(2,4-Difluoro-benzyl)-3,3,4-trimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo[3,2-b]pyridin-1-yl]-2-oxo-ethyl}-2-methyl-5-((R)-3-methyl-morpholin-4-ylmethyl)-piperazine-1-carboxylicacid tert-butyl ester

(2R,5S)-4-{2-[6-(2,4-Difluoro-benzyl)-3,3-dimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo[3,2-b]pyridin-1-yl]-2-oxo-ethyl}-2-methyl-5-((R)-3-methyl-morpholin-4-ylmethyl)-piperazine-1-carboxylicacid tert-butyl ester (670 mg, 1.04 mmol) was dissolved in THF (20 mL).Lithium tert-butoxide (170 mg, 2.08 mmol) was added, followed byiodomethane (0.16 mL, 2.60 mmol). The reaction was stirred overnight atroom temperature before being partitioned between water (30 mL) andEtOAc (2×30 mL). The organic fractions were washed with brine, driedover magnesium sulfate and concentrated. The residue was purified bycolumn chromatography, eluting with 0-10% MeOH in DCM to give the titlecompound (350 mg). MS: [M+H]⁺=658.

The following compound was prepared in an analogous method toPreparation 21:

21A:(2R,5S)-4-[2-(6-Butyl-3,3,4-trimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo[3,2-b]pyridin-1-yl)-2-oxo-ethyl]-5-((3R,5R)-3,5-dimethyl-morpholin-4-ylmethyl)-2-methyl-piperazine-1-carboxylicacid tert-butyl ester, MS: [M+H]⁺=602.

Preparation 22: tert-Butyl6-[(4-fluorophenyl)methyl]-3,3-dimethyl-4-oxy-1H,2H,3H-pyrrolo[3,2-b]pyridine-1-carboxylate

To a stirred solution of tert-butyl6-[(4-fluorophenyl)methyl]-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridine-1-carboxylate(which may be prepared as described in Preparation 15A) (3.88 g, 10.9mmol) in DCM (30 mL) at ambient temperature was added, portionwise over0.1 h, 3-chloroperbenzoic acid (77%, 2.7 g, 12.0 mmol). The mixture wasstirred for 3 h then partitioned between saturated aqueous NaHCO₃ (150mL) and DCM (3×30 mL). Combined organic extracts were dried (Na₂SO₄) andevaporated in vacuo. Residue was crystallised from ether −petrol to givethe title compound (2.62 g). 1H NMR (400 MHz, Me-d3-OD): 7.74 (1H, s),7.35-7.24 (2H, m), 7.13-7.02 (2H, m), 3.96 (2H, s), 3.79 (2H, s), 1.57(6H, s), 1.53 (9H, s).

Preparation 23: tert-Butyl6-[(4-fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridine-1-carboxylate

A mixture of tert-butyl6-[(4-fluorophenyl)methyl]-3,3-dimethyl-4-oxy-1H,2H,3H-pyrrolo[3,2-b]pyridine-1-carboxylate(which may be prepared as described in Preparation 22) (0.6 g, 1.6 mmol)and acetic anhydride (4 mL) was heated at 105° C. for 2 h then at 140°C. for 3 h, cooled then resulting solution poured into ice-water (˜100g). Resulting colourless solid was collected by filtration, then wassuspended in methanol (15 mL). Aqueous NaOH (1 M, 1.8 mL) was added andmixture stirred for 0.25 h. The solution was concentrated to 12 mL invacuo then diluted with water (20 mL) and resulting solid collected byfiltration to give the title compound (0.6 g). MS: [M+H]⁺=373.

The following compound was prepared in a similar manner to Preparation23:

23A: tert-Butyl6-[(2-fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridine-1-carboxylate

Preparation 24:6-[(4-Fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridine

A mixture of tert-butyl6-[(4-fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridine-1-carboxylate(which may be prepared as described in Preparation 23) (0.6 g, 1.6mmol), methanol (20 mL) and 5 M aqueous HCl (20 mL) was heated at refluxfor 16 h, cooled then treated with water. Resulting solid was collectedby filtration to give the title compound (0.255 g). MS: [M+H]⁺=273.

Preparation 25:1-[5-Bromo-6-(3-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone

To a solution of5-bromo-6-(3-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine(which may be prepared as described in Preparation 16A) (4.5 g, 13.43mmol) in toluene (50 mL) was added acetyl chloride (1.05 mL, 14.78 mmol)and the reaction mixture was stirred at room temperature overnight.Saturated NaHCO₃ solution (50 mL) was added and the product wasextracted with EtOAc (2×40 mL). The organic phase was washed with brine,dried, filtered and the solvent evaporated to afford the title compound(4.99 g). MS: [M+H]⁺=377.

The following compound was prepared in a similar manner to thatdescribed in Preparation 25:

25A:1-[5-Bromo-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone,MS: [M+H]⁺=377.

Preparation 26:1-[6-(3-Fluoro-benzyl)-3,3,5-trimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone

To a degassed solution of1-[5-bromo-6-(3-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 25) (4.9 g, 13.0mmol), LiBr (3.35 g, 39.0 mmol) and[1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II)dichloride (180 mg, 0.26 mmol) in THF (30 mL) and NMP (30 mL) was addedmethylzinc chloride (2M in THF, 10 mL, 20 mmol) and the reaction mixturewas stirred at room temperature overnight. The reaction mixture waspoured into water (20 mL) and 5% aqueous citric acid (3 mL) and theproduct was extracted with toluene—EtOAc (1:1, 2×40 mL). The organicphase was washed with brine, dried, filtered and the solvent evaporatedto afford the title compound (4.05 g). MS: [M+H]⁺=313.

Preparation 27:1-[6-(3-Fluoro-benzyl)-3,3,5-trimethyl-4-oxy-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone

To a solution of1-[6-(3-fluoro-benzyl)-3,3,5-trimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 26) (4.05 g, 13.0mmol) in DCM (50 mL) was added m-chloro-perbenzoic acid (77%, 4.4 g,19.5 mmol) in small portions and the reaction mixture was stirred atroom temperature for 2 h. Na₂S₂O₃ (10%, 50 mL) was added and stirred for30 mins. The product was extracted with DCM (3×40 mL), the combinedorganic layers were washed with 1M NaOH, dried, filtered and the solventevaporated to afford the title compound (4.22 g). MS: [M+H]⁺=329.

Preparation 28: Acetic acid1-acetyl-6-(3-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-ylmethylester

A solution of1-[6-(3-fluoro-benzyl)-3,3,5-trimethyl-4-oxy-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 27) (4.22 g, 12.86mmol) in acetic anhydride (25 mL) was heated at 110° C. for 2 h. Thereaction mixture was cooled, poured onto ice and stirred for 2 h. Themixture was neutralized with Na₂CO₃ and the product extracted with DCM(3×30 mL). The organic phase was dried, filtered and the solventevaporated. The crude product was purified on Silica, eluted withpetrol—EtOAc 0-50% to afford the title compound (3.49 g). MS:[M+H]⁺=371.

Preparation 29:[6-(3-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-methanol

A solution of acetic acid1-acetyl-6-(3-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-ylmethylester (which may be prepared as described in Preparation 28) (3.49 g,9.43 mmol) and NaOH (6.0 g, 150 mmol) in EtOH (60 mL) and water (60 mL)was heated at reflux overnight. The EtOH was evaporated, the pH wasadjusted to pH=8 with 5 M HCl and the product was extracted with DCM(3×30 mL). The organic phase was dried, filtered and the solventevaporated. The crude product was purified on Silica, eluted withpetrol—EtOAc 0-100% to afford the title compound (2.04 g). MS:[M+H]⁺=287.

The following compounds were prepared using a similar sequence to thatdescribed in Preparations 25-29 inclusive:

[6-(2-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-methanol,[M+H]⁺=287.

[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-methanol,[M+H]⁺=287.

[6-(2,4-Difluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-methanol,¹H NMR (400 MHz, Me-d3-OD): 7.22-7.12 (1H, m), 7.00-6.82 (2H, m),6.67-6.59 (1H, m), 4.72-4.61 (2H, m), 4.04 (2H, s), 1.34 (6H, s).

Preparation 30:1-[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone

To a solution of6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine(which may be prepared as described in Preparation 15) (10.1 g, 39 mmol)in MeCN (130 mL), at ˜10° C., was added acetyl chloride (3.6 mL, 51mmol). The mixture was stirred overnight at RT and then evaporated invacuo. The residue was partitioned between CH₂Cl₂ and 1N aqueous NaOH.CH₂Cl₂ layer was dried (MgSO₄) and evaporated to give the title compound(12.3 g) as a crystalline solid. MS: m/z=299 (M+H+)+.

Preparation 31:1-[6-(4-Fluoro-benzyl)-3,3-dimethyl-4-oxy-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone

1-[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 30) (12.2 g, 41 mmol)and mCPBA (77%, 12 g, ˜53 mmol) were dissolved in CH₂Cl₂ (150 mL) andstirred for 3 hours. 20% aqueous Na₂S₂O₃ was then added and the mixturestirred for 25 minutes. The aqueous layer was extracted with a furtherbatch of CH₂Cl₂ and then the combined CH₂Cl₂ layers were washed with2×1N aqueous NaOH. The organic layer was dried (MgSO₄) and evaporated invacuo to give the title compound (12 g) as a yellow crystalline solid.MS: m/z=315 (M+H+)+.

Preparation 32: Acetic acid1-acetyl-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-ylester

1-[6-(4-Fluoro-benzyl)-3,3-dimethyl-4-oxy-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 31) (11.55 g, 37mmol) was heated in Ac₂O (70 mL) for 5 hours. The mixture was thencooled and poured into ice/water (500 g). The mixture was stirred for 1hour and the resulting precipitate collected by filtration to give thetitle compound (12.1 g, 92%) as a grey solid. MS: m/z=357 (M+H+)+.

Preparation 33:1-Acetyl-6-(4-fluoro-benzyl)-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one

Acetic acid1-acetyl-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-ylester (which may be prepared as described in Preparation 32) (6 g, 19mmol) was suspended in EtOH (60 mL) and treated with 2N aqueous NaOH (42mL). The mixture was stirred overnight and then acidified with 5Naqueous HCl. The product was extracted with CH₂Cl₂ and the organic layerdried (MgSO₄). Purification by SiO2 chromatography (eluting with 50-100%EtOAc/hexanes) gave a yellow solid. This was triturated with toluene andthe solid collected to give the title compound (2.4 g, 44%). MS: m/z=315(M+H+)+.

The following compounds were prepared in a similar manner to thatdescribed in Preparations 30-33:

1-Acetyl-6-(2,4-difluoro-benzyl)-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=333.

1-Acetyl-6-butyl-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=263.

Preparation 34:1-Acetyl-6-(4-fluoro-benzyl)-3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one

To a mixture of1-acetyl-6-(4-fluoro-benzyl)-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one(which may be prepared as described in Preparation 33) (3.1 g, 9.9 mmol)and K2CO3 (2.7 g, 20 mmol) in DMF (30 mL), at 0° C., was addediodomethane (0.74 mL, 11.9 mmol). The mixture was allowed to stir at RTfor 5 h after which the mixture was partitioned between EtOAc and water.The EtOAc layer was washed with brine and dried (MgSO₄). Purification bySiO₂ chromatography (eluting with 0-10% MeOH/EtOAc) gave the titlecompound (960 mg, 29%) as a colourless crystalline solid. MS:[M+H]⁺=329.

Preparation 35:6-(4-Fluoro-benzyl)-3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one

1-Acetyl-6-(4-fluoro-benzyl)-3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one(which may be prepared as described in Preparation 34) (960 mg, 2.9mmol) was dissolved in a mixture of EtOH (10 mL) and 5N aqueous HCl (10mL) and heated at 95° C. for 1 hour under N₂. The mixture was thencooled and concentrated in vacuo. Ice and conc. aqueous NH₃ were addedand the resulting aqueous mixture was extracted with CH₂Cl₂. The CH₂Cl₂solution was dried (MgSO₄) and evaporated to give the title compoundwhich was used immediately. MS: [M+H]⁺=287.

The following compound was prepared in a similar manner to thatdescribed in Preparations 30-35:

6-(2,4-Difluoro-benzyl)-3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=269.

The following compounds were prepared in a similar manner to thatdescribed in Preparation 35:

6-(4-Fluoro-benzyl)-3,3,-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=273.

6-Butyl-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one, MS:[M+H]⁺=221.

6-[(2,4-Difluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridine,MS: [M+H]⁺=291.

Preparation 36:6-(2-Fluoro-benzyl)-3,3,4-trimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo[3,2-b]pyridine-1-carboxylicacid tert-butyl ester

Prepared from tert-butyl6-[(2-fluorophenyl)methyl]-3,3-dimethyl-5-oxo-1H,2H,3H,4H,5H-pyrrolo[3,2-b]pyridine-1-carboxylateusing a similar method to that described in Preparation 34. MS:[M+H]⁺=387.

Preparation 37:6-(2-Fluoro-benzyl)-3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one

Prepared from6-(2-fluoro-benzyl)-3,3,4-trimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo[3,2-b]pyridine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation36) using a similar method to that described in Preparation 24. MS:[M+H]⁺=287.

Preparation 38:1-[6-(4-Fluoro-benzyl)-3,3-dimethyl-5-vinyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone

1-[5-Bromo-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 25A) (7.64 g, 20.27mmol), tributylvinyltin (6.22 mL, 21.28 mmol) andbis(tri-tert-butylphosphine)palladium(0) (0.104 g, 0.20 mmol) weredissolved in toluene (39 mL). After being degassed with nitrogen, thereaction was heated to 85° C. for 2 h. The reaction was concentrated invacuo and the crude product was purified by column chromatography onsilica gel (gradient elution, 0-100%, ethyl acetate/petrol 40-60° C.),to give the title compound (3.64 g). MS: [M+H]⁺=325.

Preparation 39:(RS)-1-[5-(1,2-Dihydroxy-ethyl)-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone

To1-[6-(4-fluoro-benzyl)-3,3-dimethyl-5-vinyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 38) (3.64 g, 11.23mmol) in acetone (76 mL) and water (8.5 mL) was added aqueous sodiumhydroxide (2.5 M, 13.48 mL, 11.23 mmol) and the reaction cooled to 0° C.(ice bath). Potassium permanganate (1.78 g, 11.23 mol) was added to thereaction and stirred for 1 h. The reaction was warmed to roomtemperature and stirred for 20 h. Additional potassium permanganate wasadded (1.77 g, 33.7 mmol) and after 1 h the reaction was filteredthrough celite washing with acetone and water. The filtrate wasconcentrated to give an aqueous mixture which was extracted with ethylacetate (3×). The combined organics were dried over sodium sulfate,filtered and concentrated in vacuo. The crude product was purified bycolumn chromatography on silica gel (gradient elution, 0-100%, ethylacetate/petrol 40-60° C.), to give the title compound (1.5 g). MS:[M+H]⁺=359.

Chiral Purification

(RS)-1-[5-(1,2-Dihydroxy-ethyl)-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 39) (1.5 g) waspurified by chiral preparative HPLC (ChiralPAK AD-H, heptane/ethanol),to give 39A (R orS)-1-[5-(1,2-dihydroxy-ethyl)-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(fast running isomer) (0.5 g) and 39B (R orS)-1-[5-(1,2-Dihydroxy-ethyl)-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(slower running isomer) (0.6 g).

Preparation 40:(RS)-1-[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-ethane-1,2-diol

(RS)-1-[5-(1,2-Dihydroxy-ethyl)-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 39) (0.250 mg, 0.70mmol) was dissolved in ethanol (4.37 mL) and water (4.37 mL). Sodiumhydroxide (0.447 g, 11.2 mmol) was added and the reaction was heated toreflux for 4 h. After cooling to room temperature, the reaction wasconcentrated. Water was added and the aqueous was extracted with ethylacetate (3×). The combined organic extracts were dried over sodiumsulfate, filtered and concentrated, to give the title compound, (171mg). MS: [M+H]⁺=317.

The following compounds were prepared in a similar manner to thatdescribed in Preparation 40:

40A: (R orS)-1-[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-ethane-1,2-diol,from slower eluting isomer 39B. MS: [M+H]⁺=317.

40B: (R orS)-1-[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-2-methoxy-ethanol,from faster eluting precursor, MS: [M+H]⁺=331.

40C: (R orS)-1-[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-2-methoxy-ethanol,from slower eluting precursor, MS: [M+H]⁺=331.

40D: (R orS)-2-[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-2-methoxy-ethanolfrom faster eluting precursor, MS: [M+H]⁺=331.

40E: (R orS)-2-[6-(4-Fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-2-methoxy-ethanolfrom slower eluting precursor, MS: [M+H]⁺=331.

Preparation 41: (RS)-Methanesulfonic acid2-[1-acetyl-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-2-hydroxy-ethylester

To(RS)-1-[5-(1,2-dihydroxy-ethyl)-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(which may be prepared as described in Preparation 39) (1.48 g, 4.13mol) in dichloromethane (20.7 mL) cooled to 0° C. was addedtriethylamine (0.502 g, 4.96 mmol) and methane sulfonyl chloride (0.34mL, 4.34 mmol). The reaction was warmed to room temperature and stirredfor 2 h. The reaction was poured into water and extracted with DCM (3×).The combined organic extracts were dried over sodium sulfate, filteredand concentrated in vacuo. The crude product was purified by columnchromatography on silica gel (gradient elution, 0-100%, ethylacetate/petrol 40-60° C.), to give the title compound (1.25 g) MS:[M+H]⁺=437.

Preparation 42:1-[6-(4-Fluoro-benzyl)-5-(1-hydroxy-2-methoxy-ethyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(42A) and1-[6-(4-fluoro-benzyl)-5-(2-hydroxy-1-methoxy-ethyl)-3,3-dimethyl-2,3-dihydro-pyrrolo[3,2-b]pyridin-1-yl]-ethanone(42B)

To (RS)-methanesulfonic acid2-[1-acetyl-6-(4-fluoro-benzyl)-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-5-yl]-2-hydroxy-ethylester (which may be prepared as described in Preparation 41) (1.24 g,2.84 mmol) in methanol (9.48 mL), was added sodium methoxide (25%) inmethanol (1.23 mL, 5.69 mmol). After stirring for 6 h additional sodiummethoxide (25%) in methanol (1.23 mL) was added. Mixture was stirred for18 h, then sodium methoxide (25%) in methanol (1.23 mL) was added. Afterstirring for a further 22 h, water was added and the reaction wasextracted with ethyl acetate (3×). The combined organic extracts wereconcentrated in vacuo and the crude product was purified by columnchromatography on silica gel (gradient elution, 0-100%, ethylacetate/petrol 40-60° C.), to give, separately, the two title compoundsas racemic mixtures. Chiral HPLC separation was carried out as follows:

42A: ADH column, 80/20 heptane ethanol, 0.2% DEA gave faster eluting42A1 and slower eluting 42A2

42B: LUX-2 column, 80/20 heptane ethanol, 0.2% DEA gave gave fastereluting 42B1 [1H NMR (400 MHz, Me-d3-OD): 8.18 (1H, s), 7.19 (2H, dd),7.03 (2H, t), 4.66 (1H, dd), 4.21-4.05 (2H, m), 4.05-3.82 (3H, m), 3.63(1H, dd), 3.13 (3H, s), 2.24 (3H, s), 1.42 (6H, s)] and slower eluting42 B2

Preparation 43: (6-Methoxy-4-methyl-pyridin-3-yl)-carbamic acidtert-butyl ester

To a solution of 5-amino-2-methoxy-4-picoline (5.0 g, 36.2 mmol) in THF(80 mL) and saturated aqueous Na₂CO₃ (20 mL) was addeddi-tert-butyl-dicarbonate (7.9 g, 36.2 mmol) and the reaction mixturewas stirred overnight. The reaction mixture was concentrated, extractedwith DCM, washed with brine, dried, filtered and the solvent evaporatedto afford the title compound (8.8 g). MS: [M+H]⁺=239.

Preparation 44:(5-tert-Butoxycarbonylamino-2-methoxy-pyridin-4-yl)-acetic acid

To a solution of (6-methoxy-4-methyl-pyridin-3-yl)-carbamic acidtert-butyl ester (which may be prepared as described in Preparation 43)(2.8 g, 11.9 mmol) in THF (100 mL) was added sec-butyl lithium (1.4 M incyclohexane, 28 mL, 39.3 mmol) at −78° C. The reaction mixture wasstirred for 10 mins, then CO₂ gas was bubbled into it via cannula for 1h. The reaction mixture was left to warm to room temperature, quenchedwith 2N HCl. The pH was adjusted to pH=4 with 1 N NaOH and the productextracted with EtOAc. The organic layer was washed with brine, dried,filtered and the solvent was evaporated to afford the title compound(4.4 g). MS: [M+H]⁺=283.

Preparation 45:5-Methoxy-2-oxo-2,3-dihydro-pyrrolo[2,3-c]pyridine-1-carboxylic acidtert-butyl ester

A mixture of (5-tert-butoxycarbonylamino-2-methoxy-pyridin-4-yl)-aceticacid (which may be prepared as described in Preparation 44) (3.4 g, 11.9mmol), diisopropyl-ethyl-amine (4.6 mL, 26.18 mmol), EDC (2.5 g, 13.09mmol) and HOAt (1.78 g, 13.09 mmol) in DCM (50 mL) was stirred for 3 h.The reaction mixture was washed with saturated NaHCO₃, water, brine,then dried, filtered and the solvent was evaporated. The crude productwas purified on silica, eluted with petrol—EtOAc 0-50% to afford thetitle compound (2.2 g). MS: [M+H]⁺=265.

Preparation 46:5-Methoxy-3,3-dimethyl-2-oxo-2,3-dihydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester

A mixture of5-methoxy-2-oxo-2,3-dihydro-pyrrolo[2,3-c]pyridine-1-carboxylic acidtert-butyl ester (which may be prepared as described in Preparation 45)(1.94 g, 7.35 mmol), K₂CO₃ (2.33 g, 18.57 mmol) and iodomethane (1.14mL, 18.57 mmol) in acetone (25 mL) was heated at reflux for 3 h. Thereaction mixture was cooled, the solvent evaporated, the residue waspartitioned between water and DCM. The organic phase was dried, filteredand the solvent evaporated. The crude product was purified on silica,eluted with petrol—EtOAc 0-20% to afford the title compound (1.47 g).MS: [M+H]⁺=293.

Preparation 47:6-(4-Fluoro-benzyl)-3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester

A mixture of5-methoxy-3,3-dimethyl-2-oxo-2,3-dihydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation46) (1.43 g, 4.9 mmol), NaI (1.47 g, 9.8 mmol) and 4-fluorobenzylbromide (0.67 mL, 5.4 mmol) in acetonitrile (50 mL) was heated at refluxfor 5 h, stirred at room temperature overnight and heated at refluxagain for further 6 h. The reaction mixture was cooled, poured into 10%aqueous Na₂S2O3, extracted with DCM, the organic phase was dried,filtered and the solvent evaporated. The crude product was purified onsilica, eluted with petrol—EtOAc 0-100% to afford the title compound(910 mg). MS: [M+H]⁺=387.

The following compound was prepared following an analogous procedure tothat described in Preparation 47:

47A:6-(2,4-Difluoro-benzyl)-3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester, MS: [M+H]⁺=405.

Preparation 48:6-(4-Fluoro-benzyl)-3,3-dimethyl-1,6-dihydro-3H-pyrrolo[2,3-c]pyridine-2,5-dione

A solution of6-(4-fluoro-benzyl)-3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation47) (910 mg, 2.36 mmol) in TFA (5 mL) and DCM (5 mL) was stirred for 1h. The solvent was evaporated, the residue was partitioned between DCMand saturated NaHCO₃, the organic phase was dried, the solventevaporated to afford the title compound (0.67 g). MS: [M+H]⁺=287.

Preparation 49:6-(4-Fluoro-benzyl)-3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo[2,3-c]pyridin-5-one

To a solution of6-(4-fluoro-benzyl)-3,3-dimethyl-1,6-dihydro-3H-pyrrolo[2,3-c]pyridine-2,5-dione(which may be prepared as described in Preparation 48) (526 mg, 1.84mmol) in THF (30 mL) was added a solution of BH₃.Me₂S (2M, 9.7 mL, 18.4mmol) and the mixture was heated at reflux for 3 h. Cooled, MeOH (10 mL)was added carefully and heated at reflux for 2 h. The solvent wasevaporated, the residue was partitioned between DCM and saturatedNaHCO₃. The organic phase was dried, filtered and the solvent evaporatedto give the title compound (494 mg). Used without purification. MS:[M+H]⁺=273.

The following compound was prepared following an analogous procedure tothat described in Preparation 49:

49A:6-(2,4-Difluoro-benzyl)-3,3,4-trimethyl-1,2,3,6-tetrahydro-pyrrolo[2,3-c]pyridin-5-one,MS: [M+H]⁺=305.

The following compound was prepared following an analogous procedure tothat described in Preparations 47-49 inclusive:

49B:6-(2,4-Difluoro-benzyl)-3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo[2,3-c]pyridin-5-one,MS: [M+H]⁺=291.

Preparation 50:4-Amino-1-(2-chloro-acetyl)-6-(2,4-difluoro-benzyl)-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one

To a solution of1-(2-chloro-acetyl)-6-(2,4-difluoro-benzyl)-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one(which may be prepared as described in Preparation 18D) (117 mg, 0.32mmol) in DMF (2 mL) was added potassium carbonate (88 mg, 0.64 mmol))and O-(2,4-dinitrophenyl)-hydroxylamine (95 mg, 0.48 mmol). Theresulting mixture was stirred for 2 h at room temperature. 1 M aqueoussodium hydroxide (5 mL) was added and the mixture was extracted withEtOAc (2×10 mL). The organic fractions were washed with water, driedover MgSO₄ and concentrated. The residue was purified by columnchromatography, eluting with 20-65% EtOAc in petrol to give the titlecompound (79 mg) as an orange solid. MS: [M+H]⁺=382.

The following compounds were prepared following an analogous procedureto that described in Preparation 50:

50A:4-Amino-1-(2-chloro-acetyl)-6-(4-fluoro-benzyl)-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=364.

50B:4-Amino-1-(2-chloro-acetyl)-6-butyl-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo[3,2-b]pyridin-5-one,MS: [M+H]⁺=312.

Preparation 51:4-Bromo-6-(2,4-difluoro-benzyl)-3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester

N-Bromosuccinimide (529 mg, 2.97 mmol) was added to a solution of6-(2,4-difluoro-benzyl)-3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation47A) (1.0 g, 2.47 mmol) in DMF. The solution was stirred for 1.5 hoursat 60° C. The reaction mixture was cooled to room temperature, water wasadded and the product was extracted with DCM (3×). The organic phaseswere collected, dried over Na₂SO₄, filtered and concentrated in vacuo.The residue was purified by flash chromatography to give 1.1 g of thetitle compoundas a yellow solid. MS: [M+H]⁺=484.

Preparation 52:6-(2,4-Difluoro-benzyl)-3,3,4-trimethyl-1,6-dihydro-3H-pyrrolo[2,3-c]pyridine-2,5-dione

Me₂Zn solution in heptane (1M, 5.8 mL, 5.8 mmol) was slowly added to asolution of4-bromo-6-(2,4-difluoro-benzyl)-3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation51) (935 mg, 1.93 mmol) and Pd(dppf)₂Cl₂ (282 mg, 0.38 mmol) in dioxane(10 mL). The reaction mixture was stirred at 70° C. under N₂ in a sealedvessel for 1 hour. Then a second aliquot of Me₂Zn (5.8 mL, 5.8 mmol) wasadded and the stirring was maintained for 2 hours. The reaction mixturewas cooled to room temperature, quenched with sat. NaHCO₃ and extractedwith DCM. The organic phase was dried over Na₂SO₄, filtered andconcentrated in vacuo. The crude material was purified by flashchromatography to give 200 mg of the title compoundas a yellowsemi-solid. MS: [M+H]⁺=319.

Preparation 53:5-Methoxy-3,3-dimethyl-1,3-dihydro-pyrrolo[2,3-c]pyridin-2-one

Prepared from5-methoxy-3,3-dimethyl-2-oxo-2,3-dihydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester using a similar procedure to that described inPreparation 48. MS: [M+H]⁺=193.

Preparation 54:5-Methoxy-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[2,3-c]pyridine

To a solution of5-methoxy-3,3-dimethyl-1,3-dihydro-pyrrolo[2,3-c]pyridin-2-one (whichmay be prepared as described in Preparation 53) (2.8 g, 14.6 mmol) inTHF (60 mL) was added a solution of BH₃.THF (1M, 150 mL, 150 mmol) andthe mixture was stirred at room temperature overnight. MeOH (50 mL) wasadded carefully and heated at reflux for 1 h. The solvent wasevaporated, the residue was partitioned between DCM and saturatedNaHCO₃. The organic phase was dried, filtered and the solventevaporated. The crude product was purified on silica, eluted withpetrol—EtOAc 0-60% to afford the title compound (2.27 g). MS:[M+H]⁺=179.

Preparation 55:5-Methoxy-3,3-dimethyl-2,3-dihydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester

To a solution of5-methoxy-3,3-dimethyl-2,3-dihydro-1H-pyrrolo[2,3-c]pyridine (which maybe prepared as described in Preparation 54) (534 mg, 3.0 mmol) in THF(10 mL) and saturated aqueous Na₂CO₃ (4 mL) was addeddi-tert-butyl-dicarbonate (780 mg, 3.6 mmol) and the reaction mixturewas stirred overnight, then it was diluted with water, extracted withEtOAc, washed with brine, dried, filtered and the solvent evaporated toafford the title compound (760 mg). MS: [M+H]⁺=279.

Preparation 56:6-((E)-But-2-enyl)-3,3-dimethyl-5-oxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester

A mixture of5-methoxy-3,3-dimethyl-2,3-dihydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation55) (760 mg, 2.7 mmol), NaI (410 mg, 2.7 mmol) and crotyl bromide (0.33mL, 3.24 mmol) in acetonitrile (25 mL) was heated at reflux for 5 h. Thereaction mixture was cooled, poured on 10% Na₂S₂O₃, extracted with DCM,the organic phase was dried, filtered and the solvent evaporated. Thecrude product was purified on silica, eluted with petrol—EtOAc 0-70% toafford the title compound (433 mg). MS: [M+H]⁺=319.

Preparation 57:6-Butyl-3,3-dimethyl-5-oxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester

A mixture of6-((E)-but-2-enyl)-3,3-dimethyl-5-oxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester (which may be prepared as described in Preparation56) (433 mg, 1.36 mmol) and Pd/C (10%, 100 mg) in EtOH (15 mL) washydrogenated for 1 h. The catalyst was filtered, the filtrate evaporatedand the residue was purified on silica, eluted with petrol—EtOAc 0-50%to afford the title compound (387 mg). MS: [M+H]⁺=321.

Preparation 58:6-Butyl-3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo[2,3-c]pyridin-5-one

A solution of6-butyl-3,3-dimethyl-5-oxo-2,3,5,6-tetrahydro-pyrrolo[2,3-c]pyridine-1-carboxylicacid tert-butyl ester (380 mg, 1.19 mmol) in TFA (5 mL) and DCM (5 mL)was stirred for 1 h. The solvent was evaporated, the residue waspartitioned between DCM and saturated NaHCO₃, the organic phase wasdried, the solvent evaporated and the residue was purified on silica,eluted with petrol—EtOAc 0-100% to afford the title compound (170 mg).MS: [M+H]⁺=221.

Examples 1-37

The following procedure is illustrative for the preparation of Examples1-37 listed in the table below.

A mixture of tert-butyl(2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate(0.47 g), ethyl acetate (10 mL) and HCl-dioxane (4 M; 10 mL) was stirredat 20° C. for 18 h and resulting solid was collected by filtration togive1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onedihydrochloride (Example 2, 0.43 g).

By following methods similar and/or analogous to that described above,the compounds set out in the table below were prepared from thecorresponding N-Boc protected derivatives, with any significantvariations indicated below. Precursors for the N-Boc protectedderivatives are identified (by preparation number or name) in the tablebelow. The title compounds were either isolated directly as the freebase or appropriate salt without further purification, or purified forexample using mass-directed preparative HPLC, crystallization ortrituration.

1H NMR is generated at 400 MHz, in Me-d3-OD unless indicated.

Com- [M + Eg. Structure Name Method ment 1H NMR Data H]⁺ 1

2-[(2R,5R)-2-{[(3R, 5R)-3,5- Dimethylmorpholin-4- yl]methyl}-5-methylpiperazin-1-yl]- 1-{6-[(4- fluorophenyl)methyl]-5-(hydroxymethyl)-3,3- dimethyl- 1H,2H,3H-pyrrolo [3,2-b]pyridin-1-yl}ethan-1-one dihydrochloride 10 + 18 8.53 (1H, s), 7.39-7.29 (2H, m),7.21-7.11 (2H, m), 5.01 (2H, s), 4.33-4.12 (5H, m), 4.12-3.56 (10H, m),3.38-3.34 (2H, m), 3.24-3.05 (2H, m), 1.65 (6H, s), 1.50 (3H, d), 1.39(3H, d), 1.13 (3H, d). 554 2

1-{6-[(4-Fluorophenyl) methyl]-5- (hydroxymethyl)- 3,3-dimethyl-1H,2H,3H-pyrrolo [3,2-b]pyridin-1-yl}- 2-[(2R,5R)-5-methyl- 2-{[(3R)-3-methylmorpholin-4- yl]methyl}piperazin- 1-yl]ethan-1-one dihydrochloride8 + 18 8.56 (1H, s), 7.34 (2H, dd), 7.15 (2H, dd), 5.00 (2H, s),4.33-4.19 (4H, m), 4.18-3.92 (5H, m), 3.83-3.38 (8H, m), 3.16 (4H, m),1.65 (6H, s), 1.46 (3H, d), 1.38-1.25 (3H, m). 540 3

1-{2-[(2R,5R)-2- {[(3R,5R)-3,5- Dimethylmorpholin-4- yl]methyl}-5-methylpiperazin-1- yl]acetyl}-6-[(4- fluorophenyl)methyl]- 3,3-dimethyl-1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-one dihydrochloride 10 + 18A7.96 (1H, s), 7.32-7.23 (2H, m), 7.14-7.05 (2H, m), 4.09-3.93 (4H, m),3.93-3.69 (7H, m), 3.59 (4H, m), 3.28- 3.21 (3H, m), 3.18- 3.12 (1H, m),3.12-3.02 (1H, m), 1.49 (3H, d), 1.46 (3H, s), 1.44 (3H, s), 1.35 (3H,d), 1.09 (3H, d). 540 4

1-{6-[(2-Fluorophenyl) methyl]-5- (hydroxymethyl)- 3,3-dimethyl-1H,2H,3H-pyrrolo [3,2-b]pyridin-1-yl}- 2-[(2R,5R)-5-methyl- 2-{[(3R)-3-methylmorpholin-4- yl]methyl}piperazin-1- yl]ethan-1-one dihydrochloride8 + 18B 8.59 (1H, s), 7.47-7.34 (2H, m), 7.30-7.16 (2H, m), 5.05 (2H,s), 4.40-3.90 (10H, m), 3.90-2.94 (11H, m), 1.65 (6H, s), 1.45 (3H, m),1.31-1.23 (3H, m). 540 5

6-[(4-Fluorophenyl) methyl]-3,3- dimethyl-1-{2- [(2R,5R)-5-methyl-2-{[(3R)-3- methylmorpholin-4- yl]methyl}piperazin- 1-yl]acetyl}-1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-one dihydrochloride 8 + 18A8.02 (1H, s), 7.28 (2H, dd), 7.10 (2H, t), 4.10-3.78 (9H, m), 3.74-3.54(4H, m), 3.42 (2H, s), 3.31-2.93 (6H, m), 1.46 (6H, d), 1.39 (3H, m),1.28- 1.13 (3H, m). 526 6

6-[(4-Fluorophenyl) methyl]-3,3,4- trimethyl-1-{2- [(2R,5R)-5-methyl-2-{[(3R)-3- methylmorpholin-4- yl]methyl}piperazin- 1-yl]acetyl}-1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-one dihydrochloride 8 + 18C8.06 (1H, s), 7.27 (2H, dd), 7.09 (2H, t), 4.22-3.52 (18H, m), 3.52-2.87(6H, m), 1.58 (6H, d), 1.40 (3H, m), 1.24-1.16 (3H, m). 540 7

6-[(2,4-Difluorophenyl) methyl]-1-{2- [(2R,5R)-2- {[(3R,5R)-3,5-dimethylmorpholin- 4-yl]methyl}-5- methylpiperazin-1- yl]acetyl}-3,3-dimethyl- 1H,2H,3H,4H,5H- pyrrolo[3,2-b] pyridin-5-one dihydrochloride10 + 18D 8.04 (1H, s), 7.45-7.35 (1H, m), 7.06-6.94 (2H, m), 4.27-3.91(6H, m), 3.91-3.80 (4H, m), 3.80-3.47 (6H, m), 3.21-3.03 (2H, m), 1.50(3H, d), 1.46 (6H, d), 1.35 (3H, d), 1.12 (3H, d). 558 8

6-[(2,4-Difluorophenyl) methyl]-3,3- dimethyl-1-{2- [(2R,5R)-5-methyl-2-{[(3R)-3- methylmorpholin-4- yl]methyl}piperazin- 1-yl]acetyl}-1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-one dihydrochloride 8 + 18D8.03 (1H, s), 7.44-7.35 (1H, m), 7.05-6.94 (2H, m), 4.18-3.89 (6H, m),3.89-3.76 (4H, m), 3.76-3.65 (3H, m), 3.65-3.52 (2H, m), 3.25-3.14 (2H,m), 3.05 (2H, d), 1.50-1.38 (9H, m), 1.31-1.16 (3H, m). 544 9

6-[(2-Fluorophenyl) methyl]-3,3,4- trimethyl-1-{2- [(2R,5R)-5-methyl-2-{[(3R)-3- methylmorpholin-4- yl]methyl}piperazin- 1-yl]acetyl}-1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-one dihydrochloride 8 + 18E8.09 (1H, s), 7.35 (2H, m), 7.23-7.10 (2H, m), 4.08-3.78 (9H, m),3.71-3.53 (4H, m), 3.49-3.36 (2H, m), 3.29-3.13 (4H, m), 3.04 (2H, d),1.58 (6H, d), 1.52-1.38 (3H, m), 1.35-1.18 (3H, m). 540 10

1-{2-[(2R,5R)-2- {[(3R,5R)-3,5- Dimethylmorpholin- 4-yl]methyl}-5-methylpiperazin-1- yl]acetyl}-6-[(4- fluorophenyl)methyl]-3,3,4-trimethyl- 1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-onedihydrochloride 10 + 18C 8.04 (1H, s), 7.32-7.23 (2H, m), 7.15-7.02 (2H,m), 4.10-3.52 (17H, m), 3.30-3.04 (6H, m), 1.58 (6H, d), 1.48 (3H, d),1.35 (3H, d), 1.09 (3H, d). 554 11

1-{6-[(2,4- Difluorophenyl) methyl]-5- (hydroxymethyl)- 3,3-dimethyl-1H,2H,3H-pyrrolo [3,2-b]pyridin-1-yl}- 2-[(2R,5R)-5-methyl- 2-{[(3R)-3-methylmorpholin-4- yl]methyl}piperazin- 1-yl]ethan-1-one dihydrochloride8 + 18F 8.55 (1H, s), 7.50-7.41 (1H, m), 7.13-7.01 (2H, m), 5.03 (2H,s), 4.26 (4H, m), 4.13- 3.97 (4H, m), 3.85- 3.66 (3H, m), 3.51 (5H, m),3.24 (2H, m), 3.18- 2.98 (2H, m), 1.64 (6H, s), 1.47 (3H, m), 1.39-1.28(3H, m). 558 12

6-[(2,4- Difluorophenyl) methyl]-1-{2- [(2R,5R)-2- {[(3R,5R)-3,5-dimethylmorpholin- 4-yl]methyl}-5- methylpiperazin-1- yl]acetyl}-3,3,4-trimethyl- 1H,2H,3H,4H,5H- pyrrolo[3,2-b] pyridin-5-one dihydrochloride20 7.98 (1H, s), 7.33-7.23 (1H, m), 6.93-6.78 (2H, m), 3.99-3.69 (9H,m), 3.65-3.52 (8H, m), 3.09-2.93 (2H, m), 1.46 (6H, d), 1.38 (3H, d),1.23 (3H, d), 1.01 (3H, d). 572 13

1-[5-((R or S)-1,2- Dihydroxyethyl)-6- [(4-fluorophenyl)methyl]-3,3-dimethyl- 1H,2H,3H-pyrrolo[3,2- b]pyridin-1-yl]-2-[(2R,5R)-5-methyl- 2-{[(3R)-3- methylmorpholin-4- yl]methyl}piperazin-1-yl]ethan-1-one dihydrochloride 8 + 18G then prep. HPLC, slower elutingisomer 8.56-8.41 (1H, m), 7.32 (2H, m), 7.14 (2H, t), 5.35 (1H, s), 4.24(5H, m), 4.06 (3H, m), 4.01-3.94 (1H, m), 3.91 (2H, d), 3.74 (3H, m),3.60 (2H, d), 3.43 (2H, d), 3.23 (2H, m), 3.06 (2H, s), 1.64 (6H, s),1.60- 1.20 (6H, m). 570 14

1-[5-((R or S)-1,2- Dihydroxyethyl)-6- [(4-fluorophenyl)methyl]-3,3-dimethyl- 1H,2H,3H-pyrrolo [3,2-b]pyridin-1-yl]-2-[(2R,5R)-5-methyl- 2-{[(3R)-3- methylmorpholin-4- yl]methyl}piperazin-1-yl]ethan-1-one dihydrochloride 8 + 18G then prep. HPLC, slower elutingisomer 8.51 (1H, s), 7.37-7.27 (2H, m), 7.14 (2H, t), 5.36 (1H, dd),4.32- 3.93 (9H, m), 3.91 (2H, d), 3.85-3.39 (8H, m), 3.15 (4H, m), 1.64(6H, s), 1.51-1.41 (3H, m), 1.29 (3H, s). 570 15

1-{6-[(3-Fluorophenyl) methyl]-5- (hydroxymethyl)-3,3 -dimethyl-1H,2H,3H-pyrrolo [3,2-b]pyridin-1-yl}- 2-[(2R,5R)-5-methyl- 2-{[(3R)-3-methylmorpholin-4- yl]methyl}piperazin- 1-yl]ethan-1-one dihydrochloride8 + 18H 8.62 (1H, s), 7.49-7.39 (1H, m), 7.17 (1H, d), 7.13-7.02 (2H,m), 5.01 (2H, s), 4.28 (4H, d), 4.18-3.92 (5H, m), 3.77 (3H, m), 3.60(2H, m), 3.45 (4H, m), 3.14 (2H, m), 1.67 (6H, s), 1.50-1.36 (3H, m),1.31-1.22 (3H, m). 540 16

1-{2-[(2R,5R)-2- {[(3R,5R)-3,5- Dimethylmorpholin-4- yl]methyl}-5-methylpiperazin-1-yl] acetyl}-6-[(4- fluorophenyl)methyl]- 3,3-dimethyl-1H,2H,3H,5H,6H- pyrrolo[2,3- c]pyridin-5-one dihydrochloride 10 + 18I8.34 (1H, s), 7.46-7.37 (2H, m), 7.19-7.09 (2H, m), 6.58 (1H, s), 5.42(1H, d), 5.10 (1H, d), 4.07-3.88 (7H, m), 3.83 (1H, d), 3.67-3.55 (3H,m), 3.51 (1H, dd), 3.22- 3.08 (2H, m), 1.51 (3H, d), 1.44 (6H, d), 1.36(3H, d), 1.08 (3H, d). 540 17

6-[(4-Fluorophenyl) methyl]-3,3- dimethyl-1-{2- [(2R,5R)-5-methyl-2-{[(3R)-3- methylmorpholin-4- yl]methyl}piperazin- 1-yl]acetyl}-1H,2H,3H,5H,6H- pyrrolo[2,3- c]pyridin-5-one dihydrochloride 8 + 18I8.35 (1H, s), 7.46-7.36 (2H, m), 7.18-7.08 (2H, m), 6.60 (1H, s), 5.34(1H, d), 5.19 (1H, d), 4.10-3.70 (10H, m), 3.64-3.52 (2H, m), 3.14 (4H,m), 1.43 (9H, m), 1.31- 1.09 (3H, m). 526 18

1-{2-[(2R,5R)-2- {[(2S,5R)-2,5- Dimethylmorpholin-4 -yl]methyl}-5-methylpiperazin-1-yl] acetyl}-6-[(4- fluorophenyl)methyl]-3,3,4-trimethyl- 1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-onetrihydrochloride 10A + 18C 8.04 (1H, s), 7.32-7.23 (2H, m), 7.15-7.03(2H, m), 4.06-3.51 (17H, m), 3.25-3.13 (1H, m), 3.08-2.98 (2H, m), 2.82(1H, t), 2.76-2.76 (1H, m), 1.57 (6H, d), 1.40 (3H, d), 1.29 (3H, d),1.18 (3H, d). 554 19

6-[(2,4-Difluorophenyl) methyl]-3,3,4- trimethyl-1-{2-[(2R,5R)-5-methyl-2- {[(3R)-3- methylmorpholin-4- yl]methyl}piperazin-1-yl]acetyl}- 1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-onedihydrochloride 21 8.11 (1H, s), 7.45-7.35 (1H, m), 7.04-6.94 (2H, m),4.20-3.76 (10H, m), 3.71 (3H, s), 3.64-3.53 (2H, m), 3.21 (2H, s), 3.05(2H, s), 1.58 (6H, d), 1.45- 1.18 (6H, m). 558 20

6-[(2,4- Difluorophenyl) methyl]-1-{2- [(2R,5R)-2- {[(3R,5R)-3,5-dimethylmorpholin- 4-yl]methyl}-5- methylpiperazin-1- yl]acetyl}-3,3-dimethyl- 1H,2H,3H,5H,6H- pyrrolo[2,3-c] pyridin-5-one dihydrochloride10 + 18J 8.48 (1H, s), 7.63-7.53 (1H, m), 7.12-6.98 (2H, m), 6.57 (1H,s), 5.35 (1H, d), 5.23 (1H, d), 4.17-3.93 (7H, m), 3.91-3.82 (1H, m),3.82-3.56 (8H, m), 3.27-3.11 (2H, m), 1.54 (3H, d), 1.44 (6H, d), 1.37(3H, d), 1.19 (3H, d). 558 21

1-[5-((R or S)-1,2- Dihydroxyethyl)-6- [(4-fluorophenyl)methyl]-3,3-dimethyl- 1H,2H,3H-pyrrolo [3,2-b]pyridin-1-yl]-2-[(2R,5R)-2- {[(3R,5R)-3,5- dimethylmorpholin- 4-yl]methyl}-5-methylpiperazin-1- yl]ethan-1-one dihydrochloride 10 + 18K (from slowereluting pre- cursor) 8.38 (1H, s), 7.35-7.27 (2H, m), 7.14 (2H, t),5.34-5.26 (1H, m), 4.37-3.76 (14H, m), 3.76-3.55 (5H, m), 3.25-3.05 (2H,m), 1.63-1.58 (6H, m), 1.50 (3H, d), 1.38 (3H, d), 1.10 (3H, d). 584 22

6-[(2,4- Difluorophenyl) methyl]-1-{2- [(2R,5R)-2- {[(2S,5R)-2,5-dimethylmorpholin- 4-yl]methyl}-5- methylpiperazin-1- yl]acetyl}-3,3-dimethyl- 1H,2H,3H,5H,6H- pyrrolo[2,3- c]pyridin-5-one dihydrochloride10A + 18J 8.46 (1H, s), 7.61-7.52 (1H, m), 7.11-6.98 (2H, m), 6.61 (1H,s), 5.34 (1H, d), 5.26 (1H, d), 4.18-3.89 (7H, m), 3.84-3.71 (2H, m),3.71-3.57 (2H, m), 3.53-3.36 (3H, m), 3.24 (1H, dd), 3.14-3.01 (2H, m),2.97- 2.86 (1H, m), 1.46-1.41 (9H, m), 1.31 (3H, d), 1.28 (3H, d). 55823

4-Amino-6-[(2,4- difluorophenyl)methyl]- 3,3-dimethyl-1-{2-[(2R,5R)-5-methyl- 2-{[(3R)-3- methylmorpholin-4-yl]methyl}piperazin- 1-yl]acetyl}- 1H,2H,3H,4H,5H- pyrrolo[3,2-b]pyridin-5-one dihydrochloride 8 + 50 8.06 (1H, s), 7.44-7.38 (1H, m),7.04-6.95 (2H, m), 4.14-3.73 (14H, m), 3.63-3.53 (3H, m), 3.21 (2H, d),3.10-2.98 (2H, m), 1.63 (6H, d), 1.54-1.30 (3H, m), 1.24 (3H, d). 559 24

4-Amino-6-[(4- fluorophenyl)methyl]- 3,3-dimethyl-1- {2-[(2R,5R)-5-methyl- 2-{[(3R)-3- methylmorpholin-4- yl]methyl}piperazin-1-yl]acetyl}- 1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-onedihydrochloride 8 + 50A 8.01 (1H, s), 7.32-7.24 (2H, m), 7.13-7.05 (2H,m), 4.08-3.74 (9H, m), 3.59 (8H, m), 3.22-2.97 (4H, m), 1.63 (6H, d),1.44-1.16 (6H, m). 541 25

6-[(2,4- Difluorophenyl) methyl]-1-{2- [(2R,5R)-2- {[(2S,5R)-2,5-dimethylmorpholin-4- yl]methyl}-5- methylpiperazin-1- yl]acetyl}-3,3,4-trimethyl- 1H,2H,3H,4H,5H- pyrrolo[3,2-b] pyridin-5-one dihydrochloride10 A + 18L 8.09 (1H, s), 7.44-7.35 (1H, m), 7.05-6.93 (2H, m), 4.15-3.91(5H, m), 3.91-3.55 (11H, m), 3.40-3.34 (3H, m), 3.25-3.13 (1H, m),3.13-2.97 (2H, m), 2.94-2.79 (1H, m), 1.63-1.55 (6H, m), 1.41 (3H, d),1.31 (3H, d), 1.24-1.16 (3H, m). 572 26

1-{6-[(4-Fluorophenyl) methyl]-5-((R or S)-1-hydroxy-2-methoxyethyl)-3,3- dimethyl-1H,2H,3H- pyrrolo[3,2- b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2- {[(3R)-3- methylmorpholin-4- yl]methyl}piperazin-1-yl]ethan-1-one trihydrochloride 8 + 18M 8.55 (1H, s), 7.31 (2H, dd),7.19-7.08 (2H, m), 5.47 (1H, t), 4.37- 4.18 (4H, m), 4.18- 3.91 (5H, m),3.84-3.66 (5H, m), 3.64-3.57 (1H, m), 3.56-3.35 (7H, m), 3.16 (4H, m),1.65 (6H, s), 1.50-1.25 (6H, m). 584 27

1-{6-[(4-Fluorophenyl) methyl]-5-((R or S)-2-hydroxy-1-methoxyethyl)-3,3- dimethyl-1H,2H,3H- pyrrolo[3,2- b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2- {[(3R)-3- methylmorpholin-4- yl]methyl}piperazin-1-yl]ethan-1-one trihydrochloride 8 + 18N 8.57 (1H, s), 7.32 (2H, dd),7.15 (2H, t), 5.01 (1H, t), 4.28 (4H, m), 4.19-3.85 (8H, m), 3.76 (3H,m), 3.68 (3H, s), 3.66-3.57 (1H, m), 3.56-3.42 (3H, m), 3.17 (4H, m),1.65 (6H, d), 1.45 (3H, s), 1.32 (3H, s). 584 28

1-{6-[(4-Fluorophenyl) methyl]-5-((R or S)-1-hydroxy-2-methoxyethyl)-3,3- dimethyl-1H,2H,3H- pyrrolo[3,2- b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2- {[(3R)-3- methylmorpholin-4- yl]methyl}piperazin-1-yl]ethan-1-one trihydrochloride 8 + 18O 8.46 (1H, s), 7.30 (2H, dd),7.14 (2H, t), 5.48- 5.39 (1H, m), 4.37- 4.12 (5H, m), 4.01 (4H, d),3.83-3.68 (4H, m), 3.68-3.54 (2H, m), 3.45 (3H, d), 3.37 (3H, s), 3.16(4H, s), 1.63 (6H, s), 1.43 (3H, s), 1.26 (3H, s). 584 29

1-{6-[(4-Fluorophenyl) methyl]-5-((R or S)-2-hydroxy-1-methoxyethyl)-3,3- dimethyl-1H,2H,3H- pyrrolo[3,2- b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2- {[(3R)-3- methylmorpholin-4- yl]methyl}piperazin-1-yl]ethan-1-one 8 + 18P Purified by HPLC as free base 8.17 (1H, s),7.19 (2H, dd), 7.03 (2H, t), 4.68 (1H, dd), 4.22- 4.04 (3H, m), 4.01-3.86 (3H, m), 3.77 (1H, d), 3.68 (1H, dd), 3.59 (2H, d), 3.17 (3H, s),3.16-3.11 (1H, m), 3.11-2.78 (7H, m), 2.65-2.48 (2H, m), 2.28 (1H, d),1.89 (1H, d), 1.45 (3H, s), 1.42 (3H, s), 1.07 (3H, d), 0.98 (3H, d).584 30

4-Amino-6-butyl-1- {2-[(2R,5R)-2- {[(3R,5R)-3,5- dimethylmorpholin-4-yl]methyl}-5- methylpiperazin-1- yl]acetyl}-3,3- dimethyl-1H,2H,3H,4H,5H- pyrrolo[3,2- b]pyridin-5-one dihydrochloride 10 + 50B8.38 (1H, s), 4.11-3.96 (7H, m), 3.86 (2H, m), 3.70 (4H, m), 3.66- 3.56(3H, m), 3.21 (1H, dd), 3.14-3.08 (1H, m), 2.65-2.54 (2H, m), 1.64 (6H,d), 1.63-1.57 (2H, m), 1.54 (3H, d), 1.47-1.41 (2H, m), 1.37 (3H, d),1.23 (3H, d), 0.98 (3H, t). 503 31

6-[(2,4-Difluorophenyl) methyl]-3,3,4- trimethyl-1-{2-[(2R,5R)-5-methyl-2- {[(3R)-3- methylmorpholin-4- yl]methyl}piperazin-1-yl]acetyl}- 1H,2H,3H,5H,6H- pyrrolo[2,3- c]pyridin-5-onedihydrochloride 8 + 18Q 8.38 (1H, s), 7.60-7.51 (1H, m), 7.09-6.93 (2H,m), 5.28 (1H, d), 5.20 (1H, d), 4.25- 3.88 (8H, m), 3.69 (5H, m),3.56-3.46 (1H, m), 3.41 (1H, m), 3.17 (4H, m), 2.23 (3H, s), 1.52 (6H,d), 1.41 (3H, s), 1.36-1.26 (3H, m). 558 32

6-Butyl-1-{2-[(2R,5R)- 2-{[(3R,5R)-3,5- dimethylmorpholin-4-yl]methyl}-5- methylpiperazin-1- yl]acetyl}-3,3- dimethyl-1H,2H,3H,4H,5H- pyrrolo[3,2-b] pyridin-5-one dihydrochloride 10 + 18R8.58 (1H, s), 4.18 (2H, s), 4.16-4.06 (3H, m), 4.06-3.96 (2H, m), 3.91(1H, d), 3.80 (1H, s), 3.76-3.56 (3H, m), 3.45-3.36 (2H, m), 3.27-3.01(2H, m), 2.75-2.59 (2H, m), 1.72-1.58 (2H, m), 1.55 (3H, d), 1.52 (6H,s), 1.47-1.41 (2H, m), 1.38 (3H, d), 1.27- 1.24 (3H, m), 1.00 (3H, t).488 33

6-Butyl-1-{2-[(2R,5R)- 2-{[(3R,5R)-3,5- dimethylmorpholin-4-yl]methyl}-5- methylpiperazin-1-yl] acetyl}-3,3,4- trimethyl-1H,2H,3H,4H,5H- pyrrolo[3,2-b]pyridin- 5-one dihydrochloride 21A8.46-8.38 (1H, m), 4.22- 4.04 (5H, m), 4.04-3.94 (3H, m), 3.94- 3.84(2H, m), 3.72 (2H, m), 3.69 (2H, s), 3.68 (3H, s), 3.66- 3.60 (2H, m),3.24-3.12 (2H, m), 2.69-2.49 (2H, m), 1.59 (6H, d), 1.54 (3H, d), 1.49-1.39 (4H, m), 1.38 (3H, d), 1.27-1.16 (3H, m), 1.01-0.95 (3H, m). 502 34

1-[6-Butyl-5- (hydroxymethyl)-3,3- dimethyl-1H,2H,3H- pyrrolo[3,2-b]pyridin-1-yl]-2- [(2R,5R)-2-{[(3R,5R)- 3,5-dimethylmorpholin-4-yl]methyl}-5- methylpiperazin-1- yl]ethan-1-one dihydrochloride 10 +18S 8.95 (1H, s), 4.97-4.90 (2H, m), 4.41-4.21 (3H, m), 4.21-4.12 (3H,m), 4.06-3.86 (3H, m), 3.86-3.69 (4H, m), 3.67-3.55 (1H, m), 3.47-3.37(2H, m), 3.30- 3.22 (1H, m), 3.16 (1H, dd), 2.97-2.81 (2H, m), 1.70-1.65(6H, m), 1.56 (3H, d), 1.53-1.44 (4H, m), 1.41 (3H, d), 1.32-1.29 (3H,m), 1.05-1.00 (3H, m). 502 35

6-Butyl-1-{2- [(2R,5R)-2- {[(3R,5R)-3,5- dimethylmorpholin-4-yl]methyl}-5- methylpiperazin-1- yl]acetyl}-3,3- dimethyl-1H,2H,3H,5H,6H- pyrrolo[2,3-c] pyridin-5-one dihydrochloride 10 + 18T8.50 (1H, s), 6.65 (1H, s), 4.40-3.96 (10H, m), 3.91 (2H, m), 3.82- 3.69(4H, m), 3.67- 3.49 (2H, m), 3.27-3.11 (2H, m), 1.86-1.67 (2H, m), 1.55(3H, d), 1.46 (6H, s), 1.45- 1.40 (2H, m), 1.38 (3H, d), 1.25 (3H, d),1.00 (3H, t). 488 36

2-[(2R,5R)-2- {[(3R,5R)-3,5- Dimethylmorpholin- 4-yl]methyl}-5-methylpiperazin-1- yl]-1-{6-[(4- fluorophenyl)methyl]- 5-((R or S)-2-hydroxy-1- methoxyethyl)-3,3- dimethyl-1H,2H,3H- pyrrolo[3,2-b]pyridin-1-yl} ethan-1-one 10 + 18N Puri- fied by HPLC as free base8.18 (1H, s), 7.24-7.16 (2H, m), 7.08-6.99 (2H, m), 4.69 (1H, dd),4.21-3.97 (4H, m), 3.97-3.86 (2H, m), 3.72- 3.59 (2H, m), 3.59-3.47 (2H,m), 3.30- 3.20 (2H, m), 3.16 (3H, s), 3.08-2.77 (7H, m), 2.55-2.41 (2H,m), 2.19 (1H, d), 1.43 (6H, d), 1.05 (3H, d), 0.97 (6H, d). 598 37

6-Butyl-1-{2- [(2R,5R)-2- {[(2S,5R)-2,5- dimethylmorpholin-4-yl]methyl}-5- methylpiperazin-1- yl]acetyl}-3,3- dimethyl-1H,2H,3H,4H,5H- pyrrolo[3,2-b] pyridin-5-one dihydrochloride 10A + 18R8.74 (1H, s), 4.50-4.16 (4H, m), 4.16-4.05 (4H, m), 3.95 (1H, m),3.89-3.73 (2H, m), 3.69-3.62 (2H, m), 3.57- 3.49 (2H, m), 3.22-3.05 (2H,m), 3.05- 2.80 (1H, m), 2.74 (2H, t), 1.70-1.60 (2H, m), 1.56 (6H, d),1.50-1.41 (5H, m), 1.37 (3H, d), 1.31 (3H, d), 1.01 (3H, t). 488

Example 38:1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one(Free Base)

1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onedihydrochloride (1.00 g, 1.0 eq., 1.00 wt.) (which may be prepared asdescribed in Example 2) was charged to a RB flask, dissolved in water(10.0 mL, 10.0 vol, 10.00 wt.) and stirred under nitrogen at 18 to 23°C. to give a straw coloured solution (pH=4.73, T=19.3° C.). To theaqueous solution was added ethyl acetate (10.0 mL, 10.0 vol) and thebiphasic mixture was stirred at 18 to 23° C. for 5 minutes. The layerswere separated and the aqueous layer (pH=4.58, 19.6° C.) was returned tothe flask. Sodium hydrogen carbonate (388.2 mg, 3×1.05 eq., 0.4 wt.) wasadded (cautiously) and effervescence was observed. The mixture wasstirred for 20 minutes (pH=7.51, 18.2° C.), dichloromethane (5.0 mL, 5.0vol) was added and the mixture was stirred under the same conditions,for a further 5 minutes. The layers were separated, the dichloromethanelayer was retained and the aqueous layer (pH=7.66, T=17.7° C.) wasreturned to the flask. Two further extractions with dichloromethane(2×5.0 mL, 2×5.0 vol) were performed (pH=8.25, T=18.5° C. & pH=8.47,T=18.3° C.) and the combined organic layers were dried over sodiumsulfate (1.0 g, 1.0 wt.), filtered and concentrated to dryness underreduced pressure at 40° C. (400 mbar). The concentrate was then dried at40° C. (<20 mbar) over 2 hours to give a white foam (850.2 mg, 102% th.,100% corr. for input and output w/w assays), 94.3% w/w osfb (againstTCNB), that contained ethyl acetate (3.4% w/w) and dichloromethane (0.8%w/w).

Example 39:1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate (Form A)

The free base of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one(500.0 mg, 1.0 wt) (which may be prepared as described in Example 38)was charged to a 25 mL vessel and dissolved in ethanol (1.0 mL, 2.0vol). L-(+)-Lactic acid (106.2 mg, 1.0 eq) was added and the contents ofthe vessel were stirred for 1 hour at 18 to 23° C. to give a yellowsolution. After this time, TBME (9.0 mL, 18.0 vol) was charged to thevessel and the mixture was left to stir at 18 to 23° C. The progress ofthe salt crystallisation was monitored by XRPD. The lactate saltremained in solution after stirring for 16 hours at 18 to 23° C. Afterthis time, the solution was concentrated to approximately 1/4 theoriginal volume and TBME (9.0 mL, 18.0 vol) was added to give a gummysolid and clear supernatant that changed into a finely dividedsuspension after sonication and further stirring (ca. 20 hours at 18 to23° C.). The solid was isolated by filtration, dried under a stream ofnitrogen to give 365 mg of a white solid that was identified as Form Bby XRPD. The solid was oven dried at 40 to 45° C. for 67 hours to give awhite solid (325 mg, 56% th.), 98.8% w/w oasfb (against TCNB) thatcontained TBME (1.0% w/w) and water (0.6% w/w) and was indicated as FormA by XRPD. Detailed characterising data (¹H NMR, XRPD and DSC) forExample 39 is shown in FIGS. 1-3.

Example 40:1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate (Form B)

The reaction was performed according to the procedure described forExample 39, but without oven drying to afford a white solid (529.1 mg,90% th.), 96.1% w/w oasfb (against TCNB) that contained TBME (5.9% w/w)and water (3.8% w/w) and was indicated as Form B by XRPD. An alternativeprocedure to Example 40 was also used which avoided the use of ethanol.Detailed characterising data (¹H NMR, XRPD and DSC) for Example 40 isshown in FIGS. 4-6.

Example 41:1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onesulfate (Form F)

The free base of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one(500.0 mg, 1.0 wt) (which may be prepared as described in Example 38)was charged to a 10 mL vessel and dissolved in ethanol (1.0 mL, 2.0vol). Sulfuric acid (103.2 mg, 1.0 eq.) solution in ethanol (4.0 mL, 8.0vol) was added over 10 minutes with stirring at 18 to 23° C. to give aclear gel. The contents of the vessel were stirred for 1 hour at thesame temperature, during which time the gel dissolved to give a yellowsolution. Stirring was continued for 16 hours and a white suspension wasgenerated. The progress of the salt crystallisation was monitored byXRPD. Ethanol (2.0 mL, 4.0 vol) was then added to properly mobilise thesuspension and the product was isolated by filtration and dried under astream of nitrogen to give a white solid (465.2 mg, 79% th.), 94.9% w/woasfb (against TCNB) that contained ethanol (2.9% w/w) and water (3.6%w/w) and was indicated as Form F by XRPD. Detailed characterising data(¹H NMR, XRPD and DSC) for Example 41 is shown in FIGS. 7-9.

Example 42:1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-onemesylate (Form B)

The free base of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one(500.0 mg, 1.0 wt) (which may be prepared as described in Example 38)was charged to a 25 mL vessel and dissolved in 2-propanol (2.5 mL, 5.0vol). Methanesulfonic acid (276.0 mg, 3.0 eq.) was added (small exothermwas observed) and the resulting oily-gummy mixture was stirred for 1hour at 18 to 23° C. n-Heptane (10.0 mL, 20.0 vol) was added slowly over10 minutes to give a white suspension and a small quantity of gummysolid. The progress of the salt crystallisation was monitored by XRPD.The salt did not crystallise under mild conditions (stirring at 18 to23° C. for 3 days), therefore the temperature was increased to 40 to 45°C. to give a sticky gummy solid and clear supernatant. This mixture wascooled to 18 to 23° C., mobilised with a spatula and sonicated for 20minutes to provide a white suspension that contained some gummy solids.The suspension was stirred for 20 hours at the same temperature,filtered and dried under a stream of nitrogen to give a beige solid(402.9 mg, 63% th.), 99.0% w/w oasfb (against TCNB), that contained2-propanol (2.3% w/w), n-heptane (0.2% w/w), water (1.9% w/w) and wasindicated as Form B by XRPD. Detailed characterising data (¹H NMR, XRPDand DSC) for Example 42 is shown in FIGS. 10-12.

Example 43:1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate (Form C)

First Batch

The free base of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one(10.0 g, corr.) (which may be prepared as described in Example 38) wasdissolved in isopropyl acetate (80.0 mL, 8.0 vol) to give a pale yellowsolution. Solid, anhydrous L-(+)-lactic acid (1.67 g, 1.0 eq.) wascharged in one portion to the same flask and a small quantity of gum wasevident at the base of the flask. The mixture was then stirred rapidlyto mobilise the gum and the solution spontaneously nucleated and solidwas precipitated. A specimen of solid precipitant was analysed by XRPDand was consistent with Form B. n-Heptane (12.0 vol) was added and themixture was stirred at 40° C. under nitrogen for 4 days to demonstratethat the mixture would convert into Form C, the progress of which wasmonitored by XRPD (FIG. 13). The temperature of the mixture was raisedto 55° C. and stirring was continued for 24 h to complete thetransformation (FIG. 14). The product was isolated by filtration (rapid<0.5 minute), washed with isopropyl acetate/n-heptane (2.0/3.0, v/v, 5.0vol) and dried under a stream of nitrogen for 20 h to give the titlecompound as a white powder (8.89 g, 79% th.), 91.6% w/w (osfb), Form C,m.p. 172° C.

Second Batch

The free base of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one(10.0 g, corr.) (which may be prepared as described in Example 38) wasdissolved in isopropyl acetate (60.0 mL, 6.0 vol) to give a pale yellowsolution. To which was added anhydrous L-(+)-lactic acid (1.67 g, 1.0eq.) dissolved in isopropyl acetate (10.0 mL, 1.0 vol). A line rinse ofisopropyl acetate (10.0 mL, 1.0 vol) was applied and the mixture wasstirred at 18-23° C. to give a pale yellow solution.

n-Heptane (120 mL, 12.0 vol) was added drop wise over 40 min and gum wasformed on the bottom of the flask. After stirring for 1 h 40 min theappearance of the mixture had improved but the gum was still present onthe base of the flask.

The mixture was stirred for 16 h at 18-23° C. during which time the gumhad mobilised and a granular, finely divided suspension had formed. Thesuspension was filtered under nitrogen (filtration was rapid) and thecake was sampled and analysed by XRPD (Form C). The cake was washed withisopropyl acetate/n-heptane (2.0/3.0, v/v, 5.0 vol), sampled andanalysed by XRPD (Form C) and left to pull dry on the filter under astream of nitrogen over 16 h. The product consisted of a slightlyoff-white powdery solid (11.36 g, 91% corr.) Form C, 94.3% w/w (oasfb),Form C, m.p. 172° C. and contained isopropyl acetate (1.0% ww).

Detailed characterising data (¹H NMR, XRPD, DSC and optical microscopy)for Example 43 is shown in FIGS. 15-18.

Example 44:1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate (Form C)

Step 1

To a solution of tert-butyl(2R,5S)-4-(2-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-oxoethyl)-2-methyl-5-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazine-1-carboxylate(1.0 wt) (which may be prepared as described in Preparation 19) inmethanol (10 vol) cooled to <10° C. is slowly added 4M HCl in1,4-dioxane (3 vol), followed by a line rinse of methanol (0.5 vol). Themixture is warmed to 15 to 25° C. and stirred at this temperature for atleast 12 h. The reaction mixture is then warmed to 30 to 40° C. andstirred until the reaction is deemed complete by HPLC (typically >2 h).On completion the reaction solution is concentrated to dryness at up to40° C. The residue is dissolved in purified water (8 vol) and washedwith ethyl acetate (2×4 vol). The pH of the aqueous phase is adjusted topH 12 to 13 using 4M NaOH (as required) prior to extraction with ethylacetate (3×5 vol). The combined organic phases are washed with a sodiumchloride solution (5 vol) and dried over magnesium sulfate (1.0 wt) forat least 10 minutes. The solid is removed by filtration and the filtercake washed with ethyl acetate (2×2 vol). The filtrates are concentratedon a rotary evaporator at up to 40° C., the resulting concentrate isdissolved in methyl acetate (5 vol) and the solution concentrated asabove to yield the free base of1-{6-[(4-Fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one.

Step 2

To a solution of the free base of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-one(1.0 wt) (which may be prepared as described in Step 1) in methylacetate (3 vol) is added a solution of L-(+)-lactic acid (0.085 wt) inmethyl acetate (0.75 vol). A seed slurry of1-{6-[(4-fluorophenyl)methyl]-5-(hydroxymethyl)-3,3-dimethyl-1H,2H,3H-pyrrolo[3,2-b]pyridin-1-yl}-2-[(2R,5R)-5-methyl-2-{[(3R)-3-methylmorpholin-4-yl]methyl}piperazin-1-yl]ethan-1-oneL-(+)-lactate (Form C) (0.01 wt) in methyl acetate (0.08 vol) is thencharged followed by a solution of L-(+)-lactic acid (0.085 wt) in methylacetate (0.75 vol) and a line rinse of methyl acetate (0.5 vol). Thesuspension is stirred for ca. 30 minutes prior to the addition ofn-heptane (12.0 vol) over at least 1 h maintaining 15 to 25° C. Themixture is held at 15 to 25° C. and stirred for at least 2 h. The solidis removed by filtration and the filter cake washed with 2:3 methylacetate/n-heptane (5 vol). The material is dried on the filter untilsuitable for handling and then dried in an oven at up to 80° C. untilthe methyl acetate content is 0.5% w/w, to give the title compound as anoff-white to beige solid.

Example 45: Examples of Pharmaceutical Formulations

(i) Tablet Formulation

A tablet composition containing a compound of formula (I) is prepared bymixing an appropriate amount of the compound (for example 50-250 mg)with an appropriate diluent, disintegrant, compression agent and/orglidant. One possible tablet comprises 50 mg of the compound with 197 mgof lactose (BP) as diluent, and 3 mg magnesium stearate as a lubricantand compressing to form a tablet in known manner. The compressed tabletmay be film coated.

(ii) Capsule Formulation

A capsule formulation is prepared by mixing 100-250 mg of a compound offormula (I) with an equivalent amount of lactose and filling theresulting mixture into standard hard gelatin capsules. An appropriatedisintegrant and/or glidant can be included in appropriate amounts asrequired.

(iii) Injectable Formulation I

A parenteral composition for administration by injection can be preparedby dissolving a compound of formula (I) (e.g. in a salt form) in watercontaining 10% propylene glycol to give a concentration of activecompound of 1.5% by weight. The solution is then made isotonic,sterilised by filtration or by terminal sterilisation, filled into anampoule or vial or pre-filled syringe, and sealed.

(iv) Injectable Formulation II

A parenteral composition for injection is prepared by dissolving inwater a compound of formula (I) (e.g. in salt form) (2 mg/ml) andmannitol (50 mg/ml), sterile filtering the solution or by terminalsterilisation, and filling into sealable 1 ml vials or ampoules orpre-filled syringe.

(v) Injectable Formulation III

A formulation for i.v. delivery by injection or infusion can be preparedby dissolving the compound of formula (I) (e.g. in a salt form) in waterat 20 mg/ml and then adjusted for isotonicity. The vial is then sealedand sterilised by autoclaving or filled into an ampoule or vial orpre-filled syringe, sterilised by filtration and sealed.

(vi) Injectable Formulation IV

A formulation for i.v. delivery by injection or infusion can be preparedby dissolving the compound of formula (I) (e.g. in a salt form) in watercontaining a buffer (e.g. 0.2 M acetate pH 4.6) at 20 mg/ml. The vial,ampoule or pre-filled syringe is then sealed and sterilised byautoclaving or sterilized by filtration and sealed.

(vii) Subcutaneous or Intramuscular Injection Formulation

A composition for sub-cutaneous administration is prepared by mixing acompound of the formula (I) with pharmaceutical grade corn oil to give aconcentration of 5-50 mg/ml. The composition is sterilised and filledinto a suitable container.

(viii) Lyophilised Formulation I

Aliquots of formulated compound of formula (I) are put into 50 ml vialsand lyophilized. During lyophilisation, the compositions are frozenusing a one-step freezing protocol at (−45° C.). The temperature israised to −10° C. for annealing, then lowered to freezing at −45° C.,followed by primary drying at +25° C. for approximately 3400 minutes,followed by a secondary drying with increased steps if temperature to50° C. The pressure during primary and secondary drying is set at 80millitor.

(ix) Lyophilised Formulation II

Aliquots of formulated compound of formula (I) or a salt thereof asdefined herein are put into 50 mL vials and lyophilized. Duringlyophilisation, the compositions are frozen using a one-step freezingprotocol at (−45° C.). The temperature is raised to −10° C. forannealing, then lowered to freezing at −45° C., followed by primarydrying at +25° C. for approximately 3400 minutes, followed by asecondary drying with increased steps if temperature to 50° C. Thepressure during primary and secondary drying is set at 80 millitor.

(x) Lyophilised Formulation for Use in i.v. Administration III

An aqueous buffered solution is prepared by dissolving a compound offormula (I) in a buffer. The buffered solution is filled, withfiltration to remove particulate matter, into a container (such as aType 1 glass vial) which is then partially sealed (e.g. by means of aFluorotec stopper). If the compound and formulation are sufficientlystable, the formulation is sterilised by autoclaving at 121° C. for asuitable period of time. If the formulation is not stable toautoclaving, it can be sterilised using a suitable filter and filledunder sterile conditions into sterile vials. The solution is freezedried using a suitable cycle. On completion of the freeze drying cyclethe vials are back filled with nitrogen to atmospheric pressure,stoppered and secured (e.g. with an aluminium crimp). For intravenousadministration, the freeze dried solid can be reconstituted with apharmaceutically acceptable diluent, such as 0.9% saline or 5% dextrose.The solution can be dosed as is, or can be diluted further into aninfusion bag (containing a pharmaceutically acceptable diluent, such as0.9% saline or 5% dextrose), before administration.

(xi) Active Pharmaceutical Ingredient in a Bottle

A composition for oral administration is prepared by filling a bottle orvial with a compound of formula (I). The composition is thenreconstituted with a suitable diluent for example water, fruit juice, orcommercially available vehicle such as OraSweet or Syrspend. Thereconstituted solution may be dispensed into dosing cups or oralsyringes for administration.

Biological Assays

Expression and purification of XIAP, cIAP-1 and cIAP-2 BIR3 domains Therecombinant BIR3 domain of human XIAP (residues 252-350) fused to aHis-tag, human cIAP-1 (residues 267-363) fused to a GST-tag and humancIAP-2 (residues 244-337) fused to a His-tag were overexpressed fromEscherichia coli cells grown in TB medium. Protein was isolated fromlysates using Ni-NTA affinity chromatography (XIAP/cIAP-2) orglutathione sepharase 4B affinity chromatography (cIAP-1). Affinity tagsfor XIAP and cIAP-1 were cleaved with thrombin in 25 mM HEPES pH 7.5,100 mM NaCl, 50 μM Zn(OAc)₂ and 1 mM Ca(OAc)₂ followed by purificationof BIR3 domains by size-exclusion chromatography. The His-tag wasuncleaved for cIAP-2 and the protein was not concentrated above 3 mg/mLdue to aggregation induced covalent self-oligomerization issues. Thepurified protein was stored in 25 mM Tris pH 7.5, 100 mM NaCl at −80° C.

XIAP, cIAP-1 and cIAP-2 In Vitro Competitive Displacement Binding Assays

Modified SMAC peptides and compounds were tested for their ability todisplace the fluorescent tracer from either XIAP, cIAP-1 or cIAP-2. BIR3domains of cIAP-1, cIAP-2 and XIAP were incubated with test compounds orSMAC based peptides and their respective peptide probes (Peptide ProteinResearch) in assay buffer (50 mM Hepes pH 7.5, 0.025% Tween-20, 0.01%BSA, and 1 mM DTT). Positive controls consisted of BIR3 proteins andtracer (no inhibition) and negative controls contained tracer only (100%inhibition). The samples were incubated at room temperature for 1 hr(XIAP and cIAP-2) or 3 hrs (cIAP-1) prior to being read in the BMGPherastar in Fluorescence Polarization mode (FP 485 nm, 520 nm, 520 nm).IC₅₀ values were determined from dose-response plots using nonlinearleast-squares analysis.

Final Conditions for XIAP, cIAP-1 and cIAP-2 Assays

Protein Protein Conc Peptide Probe Peptide Conc XIAP 20 nMAbuRPFK(5&6FAM)-amide 5 nM cIAP-1 4 nM AbuRPFK(5&6FAM)-amide 2 nM cIAP-220 nM AVPWK(5&6FAM)-amide 2 nM

Anti-Proliferative Activity

Inhibition of cell growth is measured using the Alamar Blue assay(Nociari, M. M, Shalev, A., Benias, P., Russo, C. Journal ofImmunological Methods 1998, 213, 157-167). The method is based on theability of viable cells to reduce resazurin to its fluorescent productresorufin. For each proliferation assay cells are plated onto 96 wellplates and allowed to recover for 16 hours prior to the addition ofinhibitor compounds (in 0.1% DMSO v/v) for a further 72 hours.

At the end of the incubation period 10% (v/v) Alamar Blue is added andincubated for a further 6 hours prior to determination of fluorescentproduct at 535 nM ex/590 nM em. The anti-proliferative activities ofcompounds of the invention can be determined by measuring the ability ofthe compounds to inhibit growth in 3 cancer cell lines:

-   -   EVSA-T (human breast carcinoma) DSMZ cat. no. ACC 433    -   MDA-MB-231 (human breast carcinoma) ECACC cat. no. 92020424    -   HCT116 (human colon carcinoma) ECACC cat. no. 91091005        (insensitive cell line used as a control for non-specific        cytoxicity)

In an assay using the cell line EVSA-T, Examples 1-34 have an EC₅₀ ofless than 0.01 μM. In particular, Examples 1-3, 5-8, 10-14, 16, 18-25,27-28 and 30-32 have an EC₅₀ of less than 0.001 μM. In an assay usingthe cell line MDA-MB-231, Examples 1-34 have an EC₅₀ of less than 0.1μM. In particular, Examples 1-8, 10-14 and 18-32 have an EC₅₀ of lessthan 0.01 μM. More particularly, Examples 7-8 have an EC₅₀ of less than0.001 μM. Data for the compounds of the invention in these assays isprovided in Table 1.

Apoptosis Induction

The table below summarises the sensitivity of a panel of nine humanmelanoma cell lines that were evaluated for apoptosis induction in thepresence of 1 ng/ml TNF-α added at the same time as 1 μM Example 2 for24 hours. A range of sensitivities were observed with 3 cell lines(SK-MEL-24, WM-266-4 and WM-115) appearing the least sensitive (<20%cells apoptotic after 24 h). The table details the percentage of totalcells positive for cleaved-caspase-3 activity after 24 hour treatmentwith 1 μM Example 2 plus 1 ng/ml TNF-α by cytometry using a fluorogeniccaspase-3 substrate (NucView488—Biotium).

Melanoma Cell % Line CleavedCaspase-3 SD SK-MEL-28 63.2 3.3 SK-MEL-552.8 2.9 SK-MEL-2 49.5 2.9 RPMI-7951 44.0 5.7 MALME-3M 29.5 2.8 A37526.5 0.2 WM-115 16.7 1.1 WM-266-4 13.2 1.3 SK-MEL-24 2.1 0.5

HEK293-XIAP-Caspase-9 Immunoprecipitation (IP) MSD Assay Protocol

Stable HEK293-XIAP-Caspase-9 cells were plated out into 96-well plates[200 μl/well at 1×10⁶ cells/mL in cultured complete medium (DMEM+10%FBS+0.5 mg/mL Geneticin (Invitrogen)] and left overnight at 37° C. torecover. Compounds were added to duplicate wells in 0.1% DMSO for 2 h at37° C. Cells were lysed in 50 μl 1×MSD lysis buffer (1% Triton X-100 in20 mM Tris.Cl (pH 7.6), 150 mM NaCl including protease inhibitors) for20 min rocking at room temperature. Streptavidin high bind MSD plate(L15SB-2) were coated with biotinylated anti-FLAG M2 antibody (SigmaF9291) at 25 μl/well with a 5 μg/mL dilution of antibody in PBS for 1 h,shaking; followed by blocking for 1 h with 150 μl 3% BSA/TBST. Celllysate (25 μl) was added to the 96-well anti-FLAG coated MSD plate andplaced on shaker for 4 h at room temperature. After washing 4 times with150 μl TBST (20 mM Tris.Cl (pH 7.6), 150 mM NaCl, 0.1% Tween-20),anti-Caspase-9 [CST #9505]diluted to 5 μl/mL in MSD blocking buffer (3%BSA/TBST) was added overnight at 4° C. After washing plates 4 times with150 μl TBST, anti-rabbit-sulfo tag (MSD cat no. R32AB-1), diluted to 2μg/mL in MSD blocking buffer, was added for 2 hours at RT. Plates werewashed 4 times with 150 μl TBST, and 150 μl/well 1×MSD read buffer(R92TC-2) added before reading each plate.

EC₅₀ values were determined from dose-response plots using nonlinearleast-squares analysis. Examples 1-37 have an EC₅₀ of less than 0.1 μM.In particular, Examples 1-13, 15, 18-25, 27-28, 30-34 and 36-37 have anEC₅₀ of less than 0.01 μM. More particularly, Examples 10, 12, 23-24 and31 have an EC₅₀ of less than 0.001 μM. Data for the compounds of theinvention in this assay is provided in Table 1.

Protocol for cIAP1 Degradation MSD Assay in MDA-MB-231 Cells

MDA-MB-231 cells were plated out into 96-well plates [200 μl/well at4×10⁵ cells/mL in cultured complete medium (DMEM+10% FBS) and leftovernight at 37° C. to recover. Compounds were added to duplicate wellsin 0.1% DMSO for 2 h at 37° C. Cells were lysed in 50 μl 1×MSD lysisbuffer (1% Triton X-100 in 20 mM Tris.Cl (pH 7.6), 150 mM NaCl includingprotease inhibitors) for 20 min rocking at room temperature.Streptavidin high bind MSD plate (L15SB-2) were coated with biotinylatedanti-cIAP1 antibody (R&D Systems cat no. AF8181-biotinylated in house)at 25 μl/well with a 5 μg/mL dilution of antibody in PBS for 1 h,shaking; followed by blocking for 1 h with 150 μl 3% BSA/TBST. Celllysate (25 μl) was added to the 96-well anti-cIAP1-coated MSD plate andplaced at 4° C. overnight. After washing 4 times with 150 μl TBST (20 mMTris.Cl (pH 7.6), 150 mM NaCl, 0.1% Tween-20), anti-cIAP1-sulfo tagdetection antibody (R&D Systems cat no. AF8181-sulfo-tagged in house),diluted to 6 μg/mL in MSD blocking buffer, was added for 2 hours at RT.Plates were washed 4 times with 150 μl TBST, and 150 μl/well 1×MSD readbuffer (R92TC-2) added before reading each plate.

EC₅₀ values were determined from dose-response plots using nonlinearleast-squares analysis. Examples 1-37 have an EC₅₀ of less than 0.01 μM.In particular, Examples 1-8, 10-14, 16, 18-27, 30-34 and 37 have an EC₅₀of less than 0.001 μM. More particularly, Example 7 has an EC₅₀ of lessthan 0.0001 μM. Data for the compounds of the invention in this assay isprovided in Table 1.

Population Patch Clamp (PPC) Assay Protocol

Inhibition of the hERG channel was measured by automated patch clampassay in CHO K1 cells, stably transfected with the hERG ion channel. PPCmeasurements were performed using an lonWorks Quattro instrument(Molecular Devices Corporation, Union City, Calif.) using a 384 wellPatchPlate (Molecular Devices Corporation) with 64 apertures per well.Each concentration of test compound was tested in duplicate wells.Amphotericin B was used to obtain electrical access to the cell interiorat a final concentration of 200 μg/mL. Human ether-à-gogo related gene(hERG) currents were measured with a prepulse to +40 mV (2 s) from theholding potential of −80 mV, followed by a step to −50 mV (2 s) toelicit the deactivating tail currents, before returning to the holdingpotential for 1 s. Compounds were incubated for 600 s between the pre-and post-compound reads. The external recording solution used was 130 mMNa Gluconate, 20 mM NaCl, 4 mM KCl, 1 mM MgCl₂, 1.8 mM CaCl₂, 10 mMHepes, 5 mM Glucose, pH to 7.3 with NaOH. All data were filtered forseal quality, seal drop, and current amplitude. The maximum currentamplitude of the 3rd pulse tail current was calculated before (Pre) andafter (Post) compound addition and the amount of block assessed bydividing the Post-compound current amplitude by the Pre-compound currentamplitude. Data generated using this assay is detailed in Table 1.

Manual Patch Clamp (MPC) Assay Protocol

Inhibition of the hERG channel was measured by Manual Patch Clamp assayin HEK293 cells stably transfected with the hERG ion channel. A HEKAEPC10 amplifier and PatchMaster software were used to collect andanalyze the data for this project. Cells were plated out onto a glasscover slip, mounted on an inverted microscope and continuously bathed incontrol solution (137 mM NaCl, 4 mM KCl, 1 mm MgCl₂, 1.8 mM CaCl₂, 10 mMHepes, 10 mM Glucose, pH7.35).

After the cells had been electronically clamped and left to equilibrate,the pulse protocol was applied. The pulse protocol involved steppingfrom a holding potential of −80 mV to +40 mV for 4 s to inactivate hERGchannels, the membrane voltage was then stepped back to −50 mV for 4 sto evoke a tail current prior to returning to the holding potential.This sequence was repeated with an inter-pulse interval of 20 s. Thevoltage protocol was applied throughout the experiment starting prior todrug (0.33% DMSO control) and after cumulative additions of ascendingcompound concentrations. Evoked peak current amplitudes werecontinuously monitored throughout the experiment.

Test compounds were applied for 5 minutes or until steady state wasreached, which ever occurred earlier, before measuring the compoundeffect. The peak tail current was measured before and after eachcompound addition. Individual cell results were normalized to theirrespective vehicle control and the results were averaged. Eachconcentration of compound was measured in duplicate. 0.1 μM Cisapridewas used as a reference inhibitor.

TABLE 1 EVSA-T HEK293-X-C9 MDA-MB-231 hERG PPC prolif IP MDA MB 231cIAP1 level (IC50 or E.g. No. (μM) (μM) (μM) (μM) PI, μM) 1 0.00024 (n =2) 0.0016 (n = 3) 0.0018 (n = 2) 0.0001 (n = 4)  40 2 0.00043 (n = 4)0.0028 (n = 5) 0.0018 (n = 4) 0.00022 (n = 7) 85% @69 (n = 2) 3 0.00023(n = 3) 0.0054 (n = 3) 0.0021 (n = 3) 0.00013 (n = 3) 39% @100 4 0.0020.0026 0.0036 0.00080 47% @30  5 0.00031 0.0053 0.0025 0.00026 42% @3006 0.00055 (n = 3) 0.0020 (n = 3) 0.0050 (n = 3) 0.00042 (n = 4) 56% @2507 0.00013 (n = 2) 0.0012 (n = 2) 0.00045 (n = 2) 0.000098 (n = 2) 33%@100 8 0.00019 0.0054 0.00075 0.00012 39% @300 9 0.0042 0.0030 0.0160.0024 50% @300 10 0.00018 0.00083 (n = 2) 0.0024 0.00018 (n = 2)  35%@62.5 11 0.00031 0.0011 0.0019 0.00020  51 12 0.00024 0.00069 0.00210.00017 41% @100 13 0.00079 0.0044 0.0036 0.00026 46% @300 14 0.000910.013 0.0050 0.00053 58% @300 15 0.0078 0.0049 0.019 0.0023 55% @100 160.00095 0.014 (n = 2) 0.018 0.00070 (n = 2) 31% @100 17 0.0026 0.028 (n= 2) 0.039 0.0050 (n = 2) 59% @300 18 0.00019 0.0012 0.0031 0.00056 44%@75  19 0.00030 (n = 2) 0.0012 (n = 2) 0.0046 (n = 2) 0.00057 (n = 2)42% @100 20 0.00049 (n = 2) 0.0063 (n = 2) 0.0040 (n = 2) 0.00013 (n =2) 44% @100 21 0.00042 0.0021 0.0015 0.00018 35% @100 22 0.00056 (n = 2)0.0039 (n = 3) 0.0044 (n = 2) 0.00025 (n = 2) 47% @100 0.00028 (n = 3)23 0.00035 0.00077 0.0016 0.00033 63% @100 24 0.00035 (n = 2) 0.00058 (n= 2) 0.0032 (n = 2) 0.00022 (n = 2) 40% @100 0.00039 (n = 3) 0.0011 (n =3) 0.0034 (n = 3) 0.00019 (n = 3) 25 0.00033 0.0027 0.0017 0.00018  47%@62.5 26 0.0012 0.012 (n = 2) 0.0079 0.00053 (n = 2)  41% @31.3 270.00084 0.0060 (n = 2) 0.0052 0.00072 (n = 3)  55% @1000 0.00074 (n = 2)0.0099 (n = 3) 0.0049 (n = 2) 0.00055 (n = 4) 28 0.00082 0.0044 0.00620.0010 45% @125 29 0.0019 0.013 0.0063 0.0017 53% @500 30 0.00069 (n =2) 0.0012 (n = 3) 0.0081 (n = 2) 0.00084 (n = 3) 230 0.0010 (n = 3)0.0014 (n = 4) 0.0087 (n = 3) 0.00079 (n = 4) 31 0.00042 0.00094 0.00460.00014 140 0.00051 (n = 2) 0.0022 (n = 3) 0.0043 (n = 2) 0.00020 (n =3) 32 0.00057 0.0015 (n = 2) 0.0063 0.00019 (n = 2) 55% @500 33 0.00140.0024 (n = 2) 0.036 0.0018 (n = 2)  33% @31.3 34 0.0034 0.0038 0.0260.0013 55 35 0.016 0.013 (n = 2) 0.13 0.0022 (n = 2)  53% @1000 360.00038 0.0026 0.0025 0.0022 420 0.00048 (n = 2) 0.0042 (n = 2) 0.0027(n = 2) 0.00061 (n = 2) 37 0.0032 0.0074 0.018 0.00055 63% @500 Example259, 0.00082 (n = 19) 0.0052 (n = 41) 0.0042 (n = 19) 0.00032 (n = 25)42% @10 (n = 2) (262 and 263) of 0.00083 (n = 21) 0.0051 (n = 43) 0.0044(n = 21) 0.00032 (n = 27) 38% @10 (n = 3) WO2012/143726

Unless indicated above the data is a result of a single experiment.Where more than one data point has been obtained, the table above showsan average (e.g. geometric or arithmetic mean) of these data points (n)to 2 significant figures.

Combination Protocol for Apoptosis

Melanoma cell lines were plated out in duplicate wells of 24-well platesat 0.5×106 cells/ml the day before treatment to allow them to adhere.After incubation of the cells with compound(s) with or without 1 ng/mlTNF-α (R&D Systems) for 24 h in a CO₂ incubator at 37° C., cells wereharvested by trypsinisation. The cell pellet from the 24-well plate wasresuspended in 100 μl FACS buffer (PBS+1% fetal bovine serum).NucView488 reagent (from Biotium) was added to a final concentration of2 μM. The plate was incubated in the dark for 30 minutes beforemeasuring fluorescent stained cells in a Guava easyCyte HT cytometer(Millipore). Cleaved caspase-3 staining was recorded in the FL1 channel,with unstained and DMSO control wells being used to set the gatedstained and unstained cell populations.

Table 2 summarises the % apoptosis increases seen in either SK-MEL-28 orA375 with the indicated combination of agents included with Example 2plus 1 ng/ml TNF-α for 24 h in culture. No increase in apoptosis wasseen over this time scale with the combination agent shown in the firstcolumn of the table alone (with or without TNF-α)—data not shown.

Table 2: Increase in percentage cells apoptotic after incubation withthe indicated combination (relative to Example 2+TNF-α alone*)

% Apoptosis % Apoptosis Fold Combination Cell Line Example 2 alone*Combination Increase Interferon-α2 SK-MEL-28 42.2 (0.1 μM) 53.9 1.3 (500u/ml) Interferon-β SK-MEL-28 42.2 (0.1 μM) 70.9 1.7 (500 u/ml)Vemurafenib SK-MEL-28 51.9 (1 μM 87.3 1.7 (1 μM) Vemurafenib A375 31.2(1 μM) 61.3 2.0 (4 μM) Trametinib A375 14.4 (0.1 μM) 52.3 3.6 (0.1 μM)

Combination Protocol for Cell Proliferation

The effect of a compound of formula (I) (Compound I) in combination withan anticancer agent (Compound II) can be assessed using the followingtechnique. Cells from human cells lines (e.g. MDA-MB-231 and EVSA-T)were seeded onto 96-well tissue culture plates at a concentration of2.5×10³, 6.0×10³, or 4.0×10³ cells/well respectively. Cells were allowedto recover for 48 hours prior to addition of compound(s) or vehiclecontrol (0.35% DMSO) as follows:

Compounds were added concurrent for 96 hours. Following a total of 96hours compound incubation, cells were fixed with ice-cold 10% (w/v)trichloroacetic acid for 1 hour on ice and then washed four times withdH₂O using a plate washer (Labsystems Wellwash Ascent) and air-dried.Cells were then stained with 0.4% (w/v) Sulforhodamine B (Sigma) in 1%acetic acid for 20 min at room temperature and then washed four timeswith 1% (v/v) acetic acid and air-dried before the addition of 10 mMTris buffer to solubilise the dye. Colourmetric product was quantifiedby reading at Abs490 nm on a Wallac Victor² plate reader (1420multilabel counter, Perkin Elmer Life Sciences). The IC₅₀ for CompoundII in the presence of varying doses of Compound I was determined.Synergy was determined when the IC₅₀ shifted down in the presence ofsub-effective doses of Compound I. Additivity was determined when theresponse to Compound II and Compound I together resulted in an effectequivalent to the sum of the two compounds individually. Antagonisticeffects were defined as those causing the IC₅₀ to shift upwards, i.e.those where the response to the two compounds was less than the sum ofthe effect of the two compounds individually.

1. A compound of formula (II), (IV), or (XXII):

wherein X is CR⁴, N or NR³; wherein when X is CR⁴, then U representsnitrogen and R⁶ represents oxo; or when X is N, then U represents carbonand R⁶ represents hydroxymethyl or —CH(OR^(x))CH₂OR^(z); or when X isNR³, then U represents carbon and R⁶ represents oxo; dashed bond (

) represents a single or double bond wherein at least two of said dashedbonds represent a double bond; R³ represents hydrogen, methyl or —NH₂;R⁴ represents hydrogen, methyl, hydroxymethyl, —NH₂ or fluorine; R⁵represents unsubstituted n-butyl or benzyl substituted on the phenylgroup by one or two fluorines; R^(x) and R^(z) independently representhydrogen or methyl; L¹ represents a suitable leaving group; P¹represents hydrogen or a suitable protecting group; and L² represents asuitable leaving group.
 2. The compound of formula (II) according toclaim
 1. 3. The compound of formula (II) according to claim 1, whereinL¹ is a halogen atom and P¹ is a tert-butyloxycarbonyl (tBoc) group. 4.The compound of formula (II) according to claim 1, wherein L¹ is achlorine atom.
 5. The compound of formula (IV) according to claim
 1. 6.The compound of formula (IV) according to claim 1, wherein L² is achlorine atom.
 7. The compound of formula (XXII) according to claim 1.8. A compound of formula (XV), (XIV), (XIII), or (IV)^(a):

wherein R⁵ represents unsubstituted n-butyl or benzyl substituted on thephenyl group by one or two fluorines; L² represents a suitable leavinggroup; and L⁶ represents fluorine, bromine or chlorine.
 9. A compoundaccording to claim 8, of formula (XIII):


10. A compound according to claim 8, of formula (IV)^(a):


11. A process for the preparation of a compound of formula (IV)^(a) asrecited in claim 10, the process comprising reacting a compound offormula (XV):

with a haloacetyl halide in MeCN followed by addition of potassiumcarbonate in methanol.
 12. A process according to claim 11 wherein thehaloacetyl halide is chloroacetyl chloride.
 13. A process for thepreparation of a compound of formula (IV)^(a) as recited in claim 10,the process comprising: reacting a compound of formula (VIII):

wherein L³, L⁴, and L⁵ each individually represent fluorine, bromine orchlorine, with a base in the presence of tetrahydrofuran andisobutronitrile in a suitable solvent; and/or reacting a compound offormula (IX):

wherein L⁴ and L⁵ each individually represent fluorine, bromine orchlorine, with (I) a borane-tetrahydrofuran complex in the presence of asuitable solvent; or (II) nickel(II) chloride hexahydrate followed byaddition of sodium borohydride; and/or performing cyclization of acompound of formula (X):

wherein L⁴ and L⁵ each individually represent fluorine, bromine orchlorine, using a suitable base and an appropriate solvent; and/orreacting a compound of formula (XI):

wherein L⁵ represents fluorine, bromine or chlorine, with a compound offormula R⁵-M, wherein M represents the residue of an organometallicspecies such that R⁵-M represents a nucleophilic organometallic reagent,said reacting comprising using lithium bromide, a catalyst, and asuitable solvent system; and/or performing halogenation of a compound offormula (XII):

thereby forming a compound of formula (XIII):

wherein L⁶ represents fluorine, bromine or chlorine; and/or replacing L⁶in a compound of formula (XIII),

wherein L⁶ represents fluorine, bromine or chlorine; with a formyl groupby performing lithiation and reaction of the compound of formula (XIII)with a suitable electrophile; and/or reducing the formyl group ofcompound (XIV):

with a suitable reducing agent; and/or reacting a compound of formula(XV):

with a haloacetyl halide.
 14. The process according to claim 13, whereinL³ and L⁴ are both fluorine.
 15. The process according to claim 13,wherein L⁵ is chlorine and/or L⁶ is bromine.
 16. The process accordingto claim 13, comprising: reacting a compound of formula (VIII):

wherein L³, L⁴, and L⁵ each individually represent fluorine, bromine orchlorine, with sodium bis(trimethylsilyl)amide in the presence oftetrahydrofuran and isobutronitrile in toluene; and/or reacting acompound of formula (IX):

wherein L⁴ and L⁵ each individually represent fluorine, bromine orchlorine, with (III) a borane-tetrahydrofuran complex in the presence oftetrahydrofuran; or (IV) nickel(II) chloride hexahydrate followed byaddition of sodium borohydride; and/or performing cyclization of acompound of formula (X):

wherein L⁴ and L⁵ each individually represent fluorine, bromine orchlorine, using potassium carbonate and NMP; and/or reacting a compoundof formula (XI):

wherein L⁵ represents fluorine, bromine or chlorine, with a compound offormula R⁵-M, wherein M represents the residue of an organometallicspecies such that R⁵-M represents a nucleophilic organometallic reagentthat is an organozinc halide, said reacting comprising using lithiumbromide,[1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II)dichloride as a catalyst, and a suitable solvent system comprisingtetrahydrofuran and NMP; and/or performing halogenation of a compound offormula (XII):

using N-bromosuccinimide in dimethylformamide, thereby forming acompound of formula (XIII):

wherein L⁶ represents bromine; and/or replacing L⁶ in a compound offormula (XIII),

wherein L⁶ represents fluorine, bromine or chlorine; with a formyl groupby reacting the compound (XIII) with MeLi in THF followed by addition oftBuLi in hexane followed by addition of dimethylformamide; and/orreducing the formyl group of compound (XIV):

by reacting the compound of formula (XIV) with sodium borohydride inmethanol; and/or reacting a compound of formula (XV):

with chloroacetyl chloride in MeCN followed by addition of potassiumcarbonate in methanol.
 17. The process according to claim 16, wherein L³and L⁴ are both fluorine.
 18. The process according to claim 16, whereinL⁵ is chlorine and/or L⁶ is bromine.